Antibody Therapeutics That Bind CD137

ABSTRACT

There is disclosed compositions and methods relating to or derived from anti-CD137 antibodies. More specifically, there is disclosed fully human antibodies that bind CD137, CD137-antibody binding fragments and derivatives of such antibodies, and CD137-binding polypeptides comprising such fragments. Further still, there is disclosed nucleic acids encoding such antibodies, antibody fragments and derivatives and polypeptides, cells comprising such polynucleotides, methods of making such antibodies, antibody fragments and derivatives and polypeptides, and methods of using such antibodies, antibody fragments and derivatives and polypeptides, including methods of treating a disease requiring either stimulation of immune responses or suppression. Diseases amenable to treatment is selected from the group consisting of cancers, autoimmune diseases and viral infections.

RELATED APPLICATION

This application claims priority to U.S. Provisional Application No.62/119,211, filed on Feb. 22, 2015, the entire contents of which areincorporated by reference in its entirety herein.

TECHNICAL FIELD

The present disclosure provides compositions and methods relating to orderived from anti-CD137 antibodies. More specifically, the presentdisclosure provides fully human antibodies that bind CD137,CD137-antibody binding fragments and derivatives of such antibodies, andCD137-binding polypeptides comprising such fragments. Further still, thepresent disclosure provides nucleic acids encoding such antibodies,antibody fragments and derivatives and polypeptides, cells comprisingsuch polynucleotides, methods of making such antibodies, antibodyfragments and derivatives and polypeptides, and methods of using suchantibodies, antibody fragments and derivatives and polypeptides,including methods of treating a disease requiring either stimulation ofimmune responses or suppression.

BACKGROUND

CD137 is a member of the tumor necrosis factor (TNF) receptor family.Its alternative names are tumor necrosis factor receptor superfamilymember 9 (TNFRSF9), 4-1BB and induced by lymphocyte activation (ILA).CD137 can be expressed by activated T cells, but to a larger extent onCD8 than on CD4 T cells. In addition, CD137 expression is found ondendritic cells, follicular dendritic cells, natural killer cells,granulocytes and cells of blood vessel walls at sites of inflammation.One characterized activity of CD137 is its costimulatory activity foractivated T cells. Crosslinking of CD137 enhances T cell proliferation,IL-2 secretion survival and cytolytic activity. Further, it can enhanceimmune activity to eliminate tumors in mice.

CD137 is a T-cell costimulatory receptor induced on TCR activation (Namet al., Curr. Cancer Drug Targets, 5:357-363 (2005); Watts et al., Annu.Rev. Immunol., 23:23-68 (2005)). In addition to its expression onactivated CD4⁺ and CD8⁺ T cells, CD137 is also expressed on CD4⁺ CD25⁺regulatory T cells, natural killer (NK) and NK-T cells, monocytes,neutrophils, and dendritic cells. Its natural ligand, CD137L, has beendescribed on antigen-presenting cells including B cells,monocyte/macrophages, and dendritic cells (Watts et al., Annu. Rev.Immunol., 23:23-68 (2005)). On interaction with its ligand, CD137 leadsto increased TCR-induced T-cell proliferation, cytokine production,functional maturation, and prolonged CD8⁺ T-cell survival (Nam et al.,Curr. Cancer Drug Targets, 5:357-363 (2005), Watts et al., Annu. Rev.Immunol., 23:23-68 (2005)).

Signaling through CD137 by either CD137L or agonistic monoclonalantibodies (mAbs) against CD137 leads to increased TCR-induced T cellproliferation, cytokine production and functional maturation, andprolonged CD8+ T cell survival. These effects result from: (1) theactivation of the NF-κB, c-Jun NH2-terminal kinase/stress-activatedprotein kinase (JNK/SAPK), and p38 mitogen-activated protein kinase(MAPK) signaling pathways, and (2) the control of anti-apoptotic andcell cycle-related gene expression. Experiments performed in both CD137and CD137L-deficient mice have additionally demonstrated the importanceof CD137 costimulation in the generation of a fully competent T cellresponse.

IL-2 and IL-15 activated NK cells express CD137, and ligation of CD137by agonistic mAbs stimulates NK cell proliferation and IFN-γ secretion,but not their cytolytic activity. Furthermore, CD137-stimulated NK cellspromote the expansion of activated T cells in vitro. In accordance withtheir costimulatory function, agonist mAbs against CD137 have been shownto promote rejection of cardiac and skin allografts, eradicateestablished tumors, broaden primary antiviral CD8+ T cell responses, andincrease T cell cytolytic potential. These studies support the view thatCD137 signaling promotes T cell function which may enhance immunityagainst tumors and infection.

Other anti-CD137 antibodies have been disclosed in U.S. 2005/0095244,issued U.S. Pat. No. 7,288,638 (such as 20H4.9-IgG4 [10C7 or BMS-663513]or 20H4.9-IgG1 [BMS-663031]); U.S. Pat. No. 6,887,673 [4E9 orBMS-554271]; U.S. Pat. Nos. 7,214,493; 6,303,121; 6,569,997; 6,905,685;6,355,476; 6,362,325 [1D8 or BMS-469492; 3H3 or BMS-469497; or 3E1];U.S. Pat. No. 6,974,863 (such as 53A2); or U.S. Pat. No. 6,210,669 (suchas 1D8, 3B8, or 3E1). Additional CD137 agonistic antibodies aredescribed in U.S. Pat. Nos. 5,928,893; 6,303,121 and 6,569,997.

These and other deficiencies in the previous antibodies are overcome bythe provision of fully human antibodies to CD137 by the presentdisclosure.

SUMMARY

The present disclosure provides a fully human anti-CD137 antibody of anIgG class that binds to a CD137 epitope with a binding affinity of atleast 10⁻⁶M, which has a heavy chain variable domain sequence that is atleast 95% identical, at least 96% identical, at least 97% identical, atleast 98% identical, or at least 99% identical, to the amino acidsequences selected from the group consisting of SEQ ID NO. 1, SEQ ID NO.3, SEQ ID NO. 5, SEQ ID NO. 6, SEQ ID NO. 7, SEQ ID NO. 9, SEQ ID NO.11, SEQ ID NO. 13, SEQ ID NO. 15, SEQ ID NO. 17, SEQ ID NO. 19, SEQ IDNO. 21, SEQ ID NO. 23, SEQ ID NO. 25, SEQ ID NO. 27, SEQ ID NO. 29, SEQID NO. 31, SEQ ID NO. 33, SEQ ID NO. 35, SEQ ID NO. 37, SEQ ID NO. 39,SEQ ID NO. 41, SEQ ID NO. 43, SEQ ID NO. 45, SEQ ID NO. 47, SEQ ID NO.49, SEQ ID NO. 51, SEQ ID NO. 53, SEQ ID NO. 55, SEQ ID NO. 57, SEQ IDNO. 59, SEQ ID NO. 61, SEQ ID NO. 63, SEQ ID NO. 65, SEQ ID NO. 67, SEQID NO. 69, SEQ ID NO. 71, SEQ ID NO. 73, SEQ ID NO. 75, SEQ ID NO. 77,SEQ ID NO. 79, SEQ ID NO. 81, SEQ ID NO. 83, SEQ ID NO. 85, SEQ ID NO.87, SEQ ID NO. 89, SEQ ID NO. 91, SEQ ID NO. 93, SEQ ID NO. 95, SEQ IDNO. 97, SEQ ID NO. 99, SEQ ID NO. 101, SEQ ID NO. 103, SEQ ID NO. 105,SEQ ID NO. 107, SEQ ID NO. 109, SEQ ID NO. 111, SEQ ID NO. 113, SEQ IDNO. 115, SEQ ID NO. 117, SEQ ID NO. 119, SEQ ID NO. 121, SEQ ID NO. 123,SEQ ID NO. 125, SEQ ID NO. 127, SEQ ID NO. 129, SEQ ID NO. 130, SEQ IDNO. 131, SEQ ID NO. 132, SEQ ID NO. 133, SEQ ID NO. 134, SEQ ID NO. 135,SEQ ID NO. 136, SEQ ID NO. 137, SEQ ID NO. 138, SEQ ID NO. 139, SEQ IDNO. 140, SEQ ID NO. 141, SEQ ID NO. 142, SEQ ID NO. 143, andcombinations thereof, and that has a light chain variable domainsequence that is at least 95% identical, at least 96% identical, atleast 97% identical, at least 98% identical, or at least 99% identical,to the amino acid sequence consisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, SEQID NO. 16, SEQ ID NO. 18, SEQ ID NO. 20, SEQ ID NO. 22, SEQ ID NO. 24,SEQ ID NO. 26, SEQ ID NO. 28, SEQ ID NO. 30, SEQ ID NO. 32, SEQ ID NO.34, SEQ ID NO. 36, SEQ ID NO. 38, SEQ ID NO. 40, SEQ ID NO. 42, SEQ IDNO. 44, SEQ ID NO. 46, SEQ ID NO. 48, SEQ ID NO. 50, SEQ ID NO. 52, SEQID NO. 54, SEQ ID NO. 56, SEQ ID NO. 58, SEQ ID NO. 60, SEQ ID NO. 62,SEQ ID NO. 64, SEQ ID NO. 66, SEQ ID NO. 68, SEQ ID NO. 70, SEQ ID NO.72, SEQ ID NO. 74, SEQ ID NO. 76, SEQ ID NO. 78, SEQ ID NO. 80, SEQ IDNO. 82, SEQ ID NO. 84, SEQ ID NO. 86, SEQ ID NO. 88, SEQ ID NO. 90, SEQID NO. 92, SEQ ID NO. 94, SEQ ID NO. 96, SEQ ID NO. 98, SEQ ID NO. 100,SEQ ID NO. 102, SEQ ID NO. 104, SEQ ID NO. 106, SEQ ID NO. 108, SEQ IDNO. 110, SEQ ID NO. 112, SEQ ID NO. 114, SEQ ID NO. 116, SEQ ID NO. 118,SEQ ID NO. 120, SEQ ID NO. 122, SEQ ID NO. 124, SEQ ID NO. 126, SEQ IDNO. 128, and combinations thereof. In one embodiment, the fully humanantibody has both a heavy chain and a light chain wherein the antibodyhas a heavy chain/light chain variable domain sequence selected from thegroup consisting of SEQ ID NO. 1/SEQ ID NO. 2 (called A1 herein), SEQ IDNO. 3/SEQ ID NO. 4 (called A4 herein), SEQ ID NO. 5/SEQ ID NO. 6 (calledA11 herein), SEQ ID NO. 7/SEQ ID NO. 8 (called B1 herein), SEQ ID NO.9/SEQ ID NO. 10 (called B3 herein), SEQ ID NO. 11/SEQ ID NO. 12 (calledB12 herein), SEQ ID NO. 13/SEQ ID NO. 14 (called C2 herein), SEQ ID NO.15/SEQ ID NO. 16 (called C3 herein), SEQ ID NO. 17/SEQ ID NO. 18 (calledC7 herein), SEQ ID NO. 19/SEQ ID NO. 20 (called C11 herein), SEQ ID NO.21/SEQ ID NO. 22 (called C12 herein), SEQ ID NO. 23/SEQ ID NO. 24(called D1 herein), SEQ ID NO. 25/SEQ ID NO. 26 (called D4 herein), SEQID NO. 27/SEQ ID NO. 28 (called D6 herein), SEQ ID NO. 29/SEQ ID NO. 30(called D7 herein), SEQ ID NO. 31/SEQ ID NO. 32 (called D8 herein), SEQID NO. 33/SEQ ID NO. 34 (called D10 herein), SEQ ID NO. 35/SEQ ID NO. 36(called E2 herein), SEQ ID NO. 37/SEQ ID NO. 38 (called E5 herein), SEQID NO. 39/SEQ ID NO. 40 (called E7 herein), SEQ ID NO. 41/SEQ ID NO. 42(called F5 herein), SEQ ID NO. 43/SEQ ID NO. 44 (called F7 herein), SEQID NO. 45/SEQ ID NO. 46 (called F11 herein), SEQ ID NO. 47/SEQ ID NO. 48(called G1 herein), SEQ ID NO. 49/SEQ ID NO. 50 (called G2 herein), SEQID NO. 51/SEQ ID NO. 52 (called G3 herein), SEQ ID NO. 53/SEQ ID NO. 54(called G5 herein), SEQ ID NO. 55/SEQ ID NO. 56 (called G6 herein), SEQID NO. 57/SEQ ID NO. 58 (called G8 herein), SEQ ID NO. 59/SEQ ID NO. 60(called G12 herein), SEQ ID NO. 61/SEQ ID NO. 62 (called H4 herein), SEQID NO. 63/SEQ ID NO. 64 (called H7 herein), SEQ ID NO. 65/SEQ ID NO. 66(called H8 herein), SEQ ID NO. 67/SEQ ID NO. 68 (called H10 herein), SEQID NO. 69/SEQ ID NO. 70 (called H11 herein), SEQ ID NO. 71/SEQ ID NO. 72(called C3sh1A1 herein), SEQ ID NO. 73/SEQ ID NO. 74 (called C3sh1A2herein), SEQ ID NO. 75/SEQ ID NO. 76 (called C3sh1A5 herein), SEQ ID NO.77/SEQ ID NO. 78 (called C3sh1A9 herein), SEQ ID NO. 79/SEQ ID NO. 80(called C3sh1B2 herein), SEQ ID NO. 81/SEQ ID NO. 82 (called C3sh1B4herein), SEQ ID NO. 83/SEQ ID NO. 84 (called C3sh1B6 herein), SEQ ID NO.85/SEQ ID NO. 86 (called C3sh1B9 herein), SEQ ID NO. 87/SEQ ID NO. 88(called C3sh1C1 herein), SEQ ID NO. 89/SEQ ID NO. 90 (called C3sh1C2herein), SEQ ID NO. 91/SEQ ID NO. 92 (called C3sh1C7 herein), SEQ ID NO.93/SEQ ID NO. 94 (called C3sh1D1 herein), SEQ ID NO. 95/SEQ ID NO. 96(called C3sh1D4 herein), SEQ ID NO. 97/SEQ ID NO. 98 (called C3sh1D6herein), SEQ ID NO. 99/SEQ ID NO. 100 (called C3sh1E2 herein), SEQ IDNO. 101/SEQ ID NO. 102 (called C3sh1E7 herein), SEQ ID NO. 103/SEQ IDNO. 104 (called C3sh1E9 herein), SEQ ID NO. 105/SEQ ID NO. 106 (calledC3sh1F1 herein), SEQ ID NO. 107/SEQ ID NO. 108 (called C3sh1F10 herein),SEQ ID NO. 109/SEQ ID NO. 110 (called C3sh1F12 herein), SEQ ID NO.111/SEQ ID NO. 112 (called C3sh1F2 herein), SEQ ID NO. 113/SEQ ID NO.114 (called C3sh1G1 herein), SEQ ID NO. 115/SEQ ID NO. 116 (calledC3sh1G11 herein), SEQ ID NO. 117/SEQ ID NO. 118 (called C3sh1G2 herein),SEQ ID NO. 119/SEQ ID NO. 120 (called C3sh1G3 herein), SEQ ID NO.121/SEQ ID NO. 122 (called C3sh1G5 herein), SEQ ID NO. 123/SEQ ID NO.124 (called C3sh1G8 herein), SEQ ID NO. 125/SEQ ID NO. 126 (calledC3sh1H10 herein), SEQ ID NO. 127/SEQ ID NO. 128 (called C3sh1H4 herein),SEQ ID NO. 129/SEQ ID NO. 28 (called MA8 herein), SEQ ID NO. 130/SEQ IDNO. 28 (called MB1 herein), SEQ ID NO. 131/SEQ ID NO. 28 (called MB3herein), SEQ ID NO. 132/SEQ ID NO. 28 (called MB10 herein), SEQ ID NO.133/SEQ ID NO. 28 (called MB12 herein), SEQ ID NO. 134/SEQ ID NO. 28(called MC8 herein), SEQ ID NO. 135/SEQ ID NO. 28 (called MD1 herein),SEQ ID NO. 136/SEQ ID NO. 28 (called MD4 herein), SEQ ID NO. 137/SEQ IDNO. 28 (called MSA11 herein), SEQ ID NO. 138/SEQ ID NO. 28 (called MSB7herein), SEQ ID NO. 139/SEQ ID NO. 28 (called MSD2 herein), SEQ ID NO.140/SEQ ID NO. 28 (called MSE3 herein), SEQ ID NO. 141/SEQ ID NO. 28(called MSE5 herein), SEQ ID NO. 142/SEQ ID NO. 28 (called MSC8 herein),SEQ ID NO. 143/SEQ ID NO. 28 (called MSH1 herein), and combinationsthereof.

In one embodiment, the invention includes an isolated anti-CD137antibody, or an antigen-binding fragment thereof, comprising a heavychain variable domain comprising complementarity determining regions(CDRs) as set forth in a heavy chain variable domain amino acid sequenceselected from the group consisting of SEQ ID Nos: 1, 3, 5, 7, 9, 11, 13,15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49,51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85,87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117,119, 121, 123, 125, 127, 129, 130, 131, 132, 133, 134, 135, 136, 137,138, 139, 140, 141, 142 and 143; and comprising a light chain variabledomain comprising CDRs as set forth in a light chain variable regionamino acid sequence selected from the group consisting of SEQ ID Nos: 2,4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40,42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76,78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108,110, 112, 114, 116, 118, 120, 122, 124, 126 and 128.

The present disclosure provides a Fab fully human anti-CD137 antibodyfragment, having a variable domain region from a heavy chain and avariable domain region from a light chain, wherein the heavy chainvariable domain sequence that is at least 95% identical, at least 96%identical, at least 97% identical, at least 98% identical, or at least99% identical, to the amino acid sequences selected from the groupconsisting of SEQ ID NO. 1, SEQ ID NO. 3, SEQ ID NO. 5, SEQ ID NO. 6,SEQ ID NO. 7, SEQ ID NO. 9, SEQ ID NO. 11, SEQ ID NO. 13, SEQ ID NO. 15,SEQ ID NO. 17, SEQ ID NO. 19, SEQ ID NO. 21, SEQ ID NO. 23, SEQ ID NO.25, SEQ ID NO. 27, SEQ ID NO. 29, SEQ ID NO. 31, SEQ ID NO. 33, SEQ IDNO. 35, SEQ ID NO. 37, SEQ ID NO. 39, SEQ ID NO. 41, SEQ ID NO. 43, SEQID NO. 45, SEQ ID NO. 47, SEQ ID NO. 49, SEQ ID NO. 51, SEQ ID NO. 53,SEQ ID NO. 55, SEQ ID NO. 57, SEQ ID NO. 59, SEQ ID NO. 61, SEQ ID NO.63, SEQ ID NO. 65, SEQ ID NO. 67, SEQ ID NO. 69, SEQ ID NO. 71, SEQ IDNO. 73, SEQ ID NO. 75, SEQ ID NO. 77, SEQ ID NO. 79, SEQ ID NO. 81, SEQID NO. 83, SEQ ID NO. 85, SEQ ID NO. 87, SEQ ID NO. 89, SEQ ID NO. 91,SEQ ID NO. 93, SEQ ID NO. 95, SEQ ID NO. 97, SEQ ID NO. 99, SEQ ID NO.101, SEQ ID NO. 103, SEQ ID NO. 105, SEQ ID NO. 107, SEQ ID NO. 109, SEQID NO. 111, SEQ ID NO. 113, SEQ ID NO. 115, SEQ ID NO. 117, SEQ ID NO.119, SEQ ID NO. 121, SEQ ID NO. 123, SEQ ID NO. 125, SEQ ID NO. 127, SEQID NO. 129, SEQ ID NO. 130, SEQ ID NO. 131, SEQ ID NO. 132, SEQ ID NO.133, SEQ ID NO. 134, SEQ ID NO. 135, SEQ ID NO. 136, SEQ ID NO. 137, SEQID NO. 138, SEQ ID NO. 139, SEQ ID NO. 140, SEQ ID NO. 141, SEQ ID NO.142, SEQ ID NO. 143, and combinations thereof, and that has a lightchain variable domain sequence that is at least 95% identical, at least96% identical, at least 97% identical, at least 98% identical, or atleast 99% identical, to the amino acid sequence consisting of SEQ ID NO.2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO.12, SEQ ID NO. 14, SEQ ID NO. 16, SEQ ID NO. 18, SEQ ID NO. 20, SEQ IDNO. 22, SEQ ID NO. 24, SEQ ID NO. 26, SEQ ID NO. 28, SEQ ID NO. 30, SEQID NO. 32, SEQ ID NO. 34, SEQ ID NO. 36, SEQ ID NO. 38, SEQ ID NO. 40,SEQ ID NO. 42, SEQ ID NO. 44, SEQ ID NO. 46, SEQ ID NO. 48, SEQ ID NO.50, SEQ ID NO. 52, SEQ ID NO. 54, SEQ ID NO. 56, SEQ ID NO. 58, SEQ IDNO. 60, SEQ ID NO. 62, SEQ ID NO. 64, SEQ ID NO. 66, SEQ ID NO. 68, SEQID NO. 70, SEQ ID NO. 72, SEQ ID NO. 74, SEQ ID NO. 76, SEQ ID NO. 78,SEQ ID NO. 80, SEQ ID NO. 82, SEQ ID NO. 84, SEQ ID NO. 86, SEQ ID NO.88, SEQ ID NO. 90, SEQ ID NO. 92, SEQ ID NO. 94, SEQ ID NO. 96, SEQ IDNO. 98, SEQ ID NO. 100, SEQ ID NO. 102, SEQ ID NO. 104, SEQ ID NO. 106,SEQ ID NO. 108, SEQ ID NO. 110, SEQ ID NO. 112, SEQ ID NO. 114, SEQ IDNO. 116, SEQ ID NO. 118, SEQ ID NO. 120, SEQ ID NO. 122, SEQ ID NO. 124,SEQ ID NO. 126, SEQ ID NO. 128, and combinations thereof.

In one embodiment, the fully human antibody Fab fragment has both aheavy chain variable domain region and a light chain variable domainregion wherein the antibody has a heavy chain/light chain variabledomain sequence selected from the group consisting SEQ ID NO. 1/SEQ IDNO. 2, SEQ ID NO. 3/SEQ ID NO. 4, SEQ ID NO. 5/SEQ ID NO. 6, SEQ ID NO.7/SEQ ID NO. 8, SEQ ID NO. 9/SEQ ID NO. 10, SEQ ID NO. 11/SEQ ID NO. 12,SEQ ID NO. 13/SEQ ID NO. 14, SEQ ID NO. 15/SEQ ID NO. 16, SEQ ID NO.17/SEQ ID NO. 18, SEQ ID NO. 19/SEQ ID NO. 20, SEQ ID NO. 21/SEQ ID NO.22, SEQ ID NO. 23/SEQ ID NO. 24, SEQ ID NO. 25/SEQ ID NO. 26, SEQ ID NO.27/SEQ ID NO. 28, SEQ ID NO. 29/SEQ ID NO. 30, SEQ ID NO. 31/SEQ ID NO.32, SEQ ID NO. 33/SEQ ID NO. 34, SEQ ID NO. 35/SEQ ID NO. 36, SEQ ID NO.37/SEQ ID NO. 38, SEQ ID NO. 39/SEQ ID NO. 40, SEQ ID NO. 41/SEQ ID NO.42, SEQ ID NO. 43/SEQ ID NO. 44, SEQ ID NO. 45/SEQ ID NO. 46, SEQ ID NO.47/SEQ ID NO. 48, SEQ ID NO. 49/SEQ ID NO. 50, SEQ ID NO. 51/SEQ ID NO.52, SEQ ID NO. 53/SEQ ID NO. 54, SEQ ID NO. 55/SEQ ID NO. 56, SEQ ID NO.57/SEQ ID NO. 58, SEQ ID NO. 59/SEQ ID NO. 60, SEQ ID NO. 61/SEQ ID NO.62, SEQ ID NO. 63/SEQ ID NO. 64, SEQ ID NO. 65/SEQ ID NO. 66, SEQ ID NO.67/SEQ ID NO. 68, SEQ ID NO. 69/SEQ ID NO. 70, SEQ ID NO. 71/SEQ ID NO.72, SEQ ID NO. 73/SEQ ID NO. 74, SEQ ID NO. 75/SEQ ID NO. 76, SEQ ID NO.77/SEQ ID NO. 78, SEQ ID NO. 79/SEQ ID NO. 80, SEQ ID NO. 81/SEQ ID NO.82, SEQ ID NO. 83/SEQ ID NO. 84, SEQ ID NO. 85/SEQ ID NO. 86, SEQ ID NO.87/SEQ ID NO. 88, SEQ ID NO. 89/SEQ ID NO. 90, SEQ ID NO. 91/SEQ ID NO.92, SEQ ID NO. 93/SEQ ID NO. 94, SEQ ID NO. 95/SEQ ID NO. 96, SEQ ID NO.97/SEQ ID NO. 98, SEQ ID NO. 99/SEQ ID NO. 100, SEQ ID NO. 101/SEQ IDNO. 102, SEQ ID NO. 103/SEQ ID NO. 104, SEQ ID NO. 105/SEQ ID NO. 106,SEQ ID NO. 107/SEQ ID NO. 108, SEQ ID NO. 109/SEQ ID NO. 110, SEQ ID NO.111/SEQ ID NO. 112, SEQ ID NO. 113/SEQ ID NO. 114, SEQ ID NO. 115/SEQ IDNO. 116, SEQ ID NO. 117/SEQ ID NO. 118, SEQ ID NO. 119/SEQ ID NO. 120,SEQ ID NO. 121/SEQ ID NO. 122, SEQ ID NO. 123/SEQ ID NO. 124, SEQ ID NO.125/SEQ ID NO. 126, SEQ ID NO. 127/SEQ ID NO. 128, SEQ ID NO. 129/SEQ IDNO. 28, SEQ ID NO. 130/SEQ ID NO. 28, SEQ ID NO. 131/SEQ ID NO. 28, SEQID NO. 132/SEQ ID NO. 28, SEQ ID NO. 133/SEQ ID NO. 28, SEQ ID NO.134/SEQ ID NO. 28, SEQ ID NO. 135/SEQ ID NO. 28, SEQ ID NO. 136/SEQ IDNO. 28, SEQ ID NO. 137/SEQ ID NO. 28, SEQ ID NO. 138/SEQ ID NO. 28, SEQID NO. 139/SEQ ID NO. 28, SEQ ID NO. 140/SEQ ID NO. 28, SEQ ID NO.141/SEQ ID NO. 28, SEQ ID NO. 142/SEQ ID NO. 28, and SEQ ID NO. 143/SEQID NO. 28.

The present disclosure provides an anti-CD137 single chain humanantibody, having a variable domain region from a heavy chain and avariable domain region from a light chain and a peptide linkerconnection the heavy chain and light chain variable domain regions,wherein the heavy chain variable domain sequence that is at least 95%identical, at least 96% identical, at least 97% identical, at least 98%identical, or at least 99% identical, to the amino acid sequencesselected from the group consisting of SEQ ID NO. 1, SEQ ID NO. 3, SEQ IDNO. 5, SEQ ID NO. 6, SEQ ID NO. 7, SEQ ID NO. 9, SEQ ID NO. 11, SEQ IDNO. 13, SEQ ID NO. 15, SEQ ID NO. 17, SEQ ID NO. 19, SEQ ID NO. 21, SEQID NO. 23, SEQ ID NO. 25, SEQ ID NO. 27, SEQ ID NO. 29, SEQ ID NO. 31,SEQ ID NO. 33, SEQ ID NO. 35, SEQ ID NO. 37, SEQ ID NO. 39, SEQ ID NO.41, SEQ ID NO. 43, SEQ ID NO. 45, SEQ ID NO. 47, SEQ ID NO. 49, SEQ IDNO. 51, SEQ ID NO. 53, SEQ ID NO. 55, SEQ ID NO. 57, SEQ ID NO. 59, SEQID NO. 61, SEQ ID NO. 63, SEQ ID NO. 65, SEQ ID NO. 67, SEQ ID NO. 69,SEQ ID NO. 71, SEQ ID NO. 73, SEQ ID NO. 75, SEQ ID NO. 77, SEQ ID NO.79, SEQ ID NO. 81, SEQ ID NO. 83, SEQ ID NO. 85, SEQ ID NO. 87, SEQ IDNO. 89, SEQ ID NO. 91, SEQ ID NO. 93, SEQ ID NO. 95, SEQ ID NO. 97, SEQID NO. 99, SEQ ID NO. 101, SEQ ID NO. 103, SEQ ID NO. 105, SEQ ID NO.107, SEQ ID NO. 109, SEQ ID NO. 111, SEQ ID NO. 113, SEQ ID NO. 115, SEQID NO. 117, SEQ ID NO. 119, SEQ ID NO. 121, SEQ ID NO. 123, SEQ ID NO.125, SEQ ID NO. 127, SEQ ID NO. 129, SEQ ID NO. 130, SEQ ID NO. 131, SEQID NO. 132, SEQ ID NO. 133, SEQ ID NO. 134, SEQ ID NO. 135, SEQ ID NO.136, SEQ ID NO. 137, SEQ ID NO. 138, SEQ ID NO. 139, SEQ ID NO. 140, SEQID NO. 141, SEQ ID NO. 142, SEQ ID NO. 143, and that has a light chainvariable domain sequence that is at least 95% identical, at least 96%identical, at least 97% identical, at least 98% identical, or at least99% identical, to the amino acid sequence consisting of SEQ ID NO. 2,SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12,SEQ ID NO. 14, SEQ ID NO. 16, SEQ ID NO. 18, SEQ ID NO. 20, SEQ ID NO.22, SEQ ID NO. 24, SEQ ID NO. 26, SEQ ID NO. 28, SEQ ID NO. 30, SEQ IDNO. 32, SEQ ID NO. 34, SEQ ID NO. 36, SEQ ID NO. 38, SEQ ID NO. 40, SEQID NO. 42, SEQ ID NO. 44, SEQ ID NO. 46, SEQ ID NO. 48, SEQ ID NO. 50,SEQ ID NO. 52, SEQ ID NO. 54, SEQ ID NO. 56, SEQ ID NO. 58, SEQ ID NO.60, SEQ ID NO. 62, SEQ ID NO. 64, SEQ ID NO. 66, SEQ ID NO. 68, SEQ IDNO. 70, SEQ ID NO. 72, SEQ ID NO. 74, SEQ ID NO. 76, SEQ ID NO. 78, SEQID NO. 80, SEQ ID NO. 82, SEQ ID NO. 84, SEQ ID NO. 86, SEQ ID NO. 88,SEQ ID NO. 90, SEQ ID NO. 92, SEQ ID NO. 94, SEQ ID NO. 96, SEQ ID NO.98, SEQ ID NO. 100, SEQ ID NO. 102, SEQ ID NO. 104, SEQ ID NO. 106, SEQID NO. 108, SEQ ID NO. 110, SEQ ID NO. 112, SEQ ID NO. 114, SEQ ID NO.116, SEQ ID NO. 118, SEQ ID NO. 120, SEQ ID NO. 122, SEQ ID NO. 124, SEQID NO. 126, SEQ ID NO. 128, and combinations thereof.

In certain embodiments, the fully human single chain antibody has both aheavy chain variable domain region and a light chain variable domainregion, wherein the single chain fully human antibody has a heavychain/light chain variable domain sequence selected from the groupconsisting of SEQ ID NO. 1/SEQ ID NO. 2, SEQ ID NO. 3/SEQ ID NO. 4, SEQID NO. 5/SEQ ID NO. 6, SEQ ID NO. 7/SEQ ID NO. 8, SEQ ID NO. 9/SEQ IDNO. 10, SEQ ID NO. 11/SEQ ID NO. 12, SEQ ID NO. 13/SEQ ID NO. 14, SEQ IDNO. 15/SEQ ID NO. 16, SEQ ID NO. 17/SEQ ID NO. 18, SEQ ID NO. 19/SEQ IDNO. 20, SEQ ID NO. 21/SEQ ID NO. 22, SEQ ID NO. 23/SEQ ID NO. 24, SEQ IDNO. 25/SEQ ID NO. 26, SEQ ID NO. 27/SEQ ID NO. 28, SEQ ID NO. 29/SEQ IDNO. 30, SEQ ID NO. 31/SEQ ID NO. 32, SEQ ID NO. 33/SEQ ID NO. 34, SEQ IDNO. 35/SEQ ID NO. 36, SEQ ID NO. 37/SEQ ID NO. 38, SEQ ID NO. 39/SEQ IDNO. 40, SEQ ID NO. 41/SEQ ID NO. 42, SEQ ID NO. 43/SEQ ID NO. 44, SEQ IDNO. 45/SEQ ID NO. 46, SEQ ID NO. 47/SEQ ID NO. 48, SEQ ID NO. 49/SEQ IDNO. 50, SEQ ID NO. 51/SEQ ID NO. 52, SEQ ID NO. 53/SEQ ID NO. 54, SEQ IDNO. 55/SEQ ID NO. 56, SEQ ID NO. 57/SEQ ID NO. 58, SEQ ID NO. 59/SEQ IDNO. 60, SEQ ID NO. 61/SEQ ID NO. 62, SEQ ID NO. 63/SEQ ID NO. 64, SEQ IDNO. 65/SEQ ID NO. 66, SEQ ID NO. 67/SEQ ID NO. 68, SEQ ID NO. 69/SEQ IDNO. 70, SEQ ID NO. 71/SEQ ID NO. 72, SEQ ID NO. 73/SEQ ID NO. 74, SEQ IDNO. 75/SEQ ID NO. 76, SEQ ID NO. 77/SEQ ID NO. 78, SEQ ID NO. 79/SEQ IDNO. 80, SEQ ID NO. 81/SEQ ID NO. 82, SEQ ID NO. 83/SEQ ID NO. 84, SEQ IDNO. 85/SEQ ID NO. 86, SEQ ID NO. 87/SEQ ID NO. 88, SEQ ID NO. 89/SEQ IDNO. 90, SEQ ID NO. 91/SEQ ID NO. 92, SEQ ID NO. 93/SEQ ID NO. 94, SEQ IDNO. 95/SEQ ID NO. 96, SEQ ID NO. 97/SEQ ID NO. 98, SEQ ID NO. 99/SEQ IDNO. 100, SEQ ID NO. 101/SEQ ID NO. 102, SEQ ID NO. 103/SEQ ID NO. 104,SEQ ID NO. 105/SEQ ID NO. 106, SEQ ID NO. 107/SEQ ID NO. 108, SEQ ID NO.109/SEQ ID NO. 110, SEQ ID NO. 111/SEQ ID NO. 112, SEQ ID NO. 113/SEQ IDNO. 114, SEQ ID NO. 115/SEQ ID NO. 116, SEQ ID NO. 117/SEQ ID NO. 118,SEQ ID NO. 119/SEQ ID NO. 120, SEQ ID NO. 121/SEQ ID NO. 122, SEQ ID NO.123/SEQ ID NO. 124, SEQ ID NO. 125/SEQ ID NO. 126, SEQ ID NO. 127/SEQ IDNO. 128, SEQ ID NO. 129/SEQ ID NO. 28, SEQ ID NO. 130/SEQ ID NO. 28, SEQID NO. 131/SEQ ID NO. 28, SEQ ID NO. 132/SEQ ID NO. 28, SEQ ID NO.133/SEQ ID NO. 28, SEQ ID NO. 134/SEQ ID NO. 28, SEQ ID NO. 135/SEQ IDNO. 28, SEQ ID NO. 136/SEQ ID NO. 28, SEQ ID NO. 137/SEQ ID NO. 28, SEQID NO. 138/SEQ ID NO. 28, SEQ ID NO. 139/SEQ ID NO. 28, SEQ ID NO.140/SEQ ID NO. 28, SEQ ID NO. 141/SEQ ID NO. 28, SEQ ID NO. 142/SEQ IDNO. 28, SEQ ID NO. 143/SEQ ID NO. 28, and combinations thereof.

The present disclosure further provides a method for treating a diseaserequiring either stimulation of immune responses or suppression,comprising administering an anti-CD137 polypeptide, wherein the fullyhuman antibody has a heavy chain variable domain sequence that is atleast 95% identical, at least 96% identical, at least 97% identical, atleast 98% identical, or at least 99% identical, to the amino acidsequences selected from the group consisting of SEQ ID NO. 1, SEQ ID NO.3, SEQ ID NO. 5, SEQ ID NO. 6, SEQ ID NO. 7, SEQ ID NO. 9, SEQ ID NO.11, SEQ ID NO. 13, SEQ ID NO. 15, SEQ ID NO. 17, SEQ ID NO. 19, SEQ IDNO. 21, SEQ ID NO. 23, SEQ ID NO. 25, SEQ ID NO. 27, SEQ ID NO. 29, SEQID NO. 31, SEQ ID NO. 33, SEQ ID NO. 35, SEQ ID NO. 37, SEQ ID NO. 39,SEQ ID NO. 41, SEQ ID NO. 43, SEQ ID NO. 45, SEQ ID NO. 47, SEQ ID NO.49, SEQ ID NO. 51, SEQ ID NO. 53, SEQ ID NO. 55, SEQ ID NO. 57, SEQ IDNO. 59, SEQ ID NO. 61, SEQ ID NO. 63, SEQ ID NO. 65, SEQ ID NO. 67, SEQID NO. 69, SEQ ID NO. 71, SEQ ID NO. 73, SEQ ID NO. 75, SEQ ID NO. 77,SEQ ID NO. 79, SEQ ID NO. 81, SEQ ID NO. 83, SEQ ID NO. 85, SEQ ID NO.87, SEQ ID NO. 89, SEQ ID NO. 91, SEQ ID NO. 93, SEQ ID NO. 95, SEQ IDNO. 97, SEQ ID NO. 99, SEQ ID NO. 101, SEQ ID NO. 103, SEQ ID NO. 105,SEQ ID NO. 107, SEQ ID NO. 109, SEQ ID NO. 111, SEQ ID NO. 113, SEQ IDNO. 115, SEQ ID NO. 117, SEQ ID NO. 119, SEQ ID NO. 121, SEQ ID NO. 123,SEQ ID NO. 125, SEQ ID NO. 127, SEQ ID NO. 129, SEQ ID NO. 130, SEQ IDNO. 131, SEQ ID NO. 132, SEQ ID NO. 133, SEQ ID NO. 134, SEQ ID NO. 135,SEQ ID NO. 136, SEQ ID NO. 137, SEQ ID NO. 138, SEQ ID NO. 139, SEQ IDNO. 140, SEQ ID NO. 141, SEQ ID NO. 142, SEQ ID NO. 143, andcombinations thereof, and that has a light chain variable domainsequence that is at least 95% identical to the amino acid consisting ofSEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10,SEQ ID NO. 12, SEQ ID NO. 14, SEQ ID NO. 16, SEQ ID NO. 18, SEQ ID NO.20, SEQ ID NO. 22, SEQ ID NO. 24, SEQ ID NO. 26, SEQ ID NO. 28, SEQ IDNO. 30, SEQ ID NO. 32, SEQ ID NO. 34, SEQ ID NO. 36, SEQ ID NO. 38, SEQID NO. 40, SEQ ID NO. 42, SEQ ID NO. 44, SEQ ID NO. 46, SEQ ID NO. 48,SEQ ID NO. 50, SEQ ID NO. 52, SEQ ID NO. 54, SEQ ID NO. 56, SEQ ID NO.58, SEQ ID NO. 60, SEQ ID NO. 62, SEQ ID NO. 64, SEQ ID NO. 66, SEQ IDNO. 68, SEQ ID NO. 70, SEQ ID NO. 72, SEQ ID NO. 74, SEQ ID NO. 76, SEQID NO. 78, SEQ ID NO. 80, SEQ ID NO. 82, SEQ ID NO. 84, SEQ ID NO. 86,SEQ ID NO. 88, SEQ ID NO. 90, SEQ ID NO. 92, SEQ ID NO. 94, SEQ ID NO.96, SEQ ID NO. 98, SEQ ID NO. 100, SEQ ID NO. 102, SEQ ID NO. 104, SEQID NO. 106, SEQ ID NO. 108, SEQ ID NO. 110, SEQ ID NO. 112, SEQ ID NO.114, SEQ ID NO. 116, SEQ ID NO. 118, SEQ ID NO. 120, SEQ ID NO. 122, SEQID NO. 124, SEQ ID NO. 126, SEQ ID NO. 128, combinations thereof;wherein the Fab fully human antibody fragment has the heavy chainvariable domain sequence that is at least 95% identical, at least 96%identical, at least 97% identical, at least 98% identical, or at least99% identical, to the amino acid sequences selected from the groupconsisting of SEQ ID NO. 1, SEQ ID NO. 3, SEQ ID NO. 5, SEQ ID NO. 6,SEQ ID NO. 7, SEQ ID NO. 9, SEQ ID NO. 11, SEQ ID NO. 13, SEQ ID NO. 15,SEQ ID NO. 17, SEQ ID NO. 19, SEQ ID NO. 21, SEQ ID NO. 23, SEQ ID NO.25, SEQ ID NO. 27, SEQ ID NO. 29, SEQ ID NO. 31, SEQ ID NO. 33, SEQ IDNO. 35, SEQ ID NO. 37, SEQ ID NO. 39, SEQ ID NO. 41, SEQ ID NO. 43, SEQID NO. 45, SEQ ID NO. 47, SEQ ID NO. 49, SEQ ID NO. 51, SEQ ID NO. 53,SEQ ID NO. 55, SEQ ID NO. 57, SEQ ID NO. 59, SEQ ID NO. 61, SEQ ID NO.63, SEQ ID NO. 65, SEQ ID NO. 67, SEQ ID NO. 69, SEQ ID NO. 71, SEQ IDNO. 73, SEQ ID NO. 75, SEQ ID NO. 77, SEQ ID NO. 79, SEQ ID NO. 81, SEQID NO. 83, SEQ ID NO. 85, SEQ ID NO. 87, SEQ ID NO. 89, SEQ ID NO. 91,SEQ ID NO. 93, SEQ ID NO. 95, SEQ ID NO. 97, SEQ ID NO. 99, SEQ ID NO.101, SEQ ID NO. 103, SEQ ID NO. 105, SEQ ID NO. 107, SEQ ID NO. 109, SEQID NO. 111, SEQ ID NO. 113, SEQ ID NO. 115, SEQ ID NO. 117, SEQ ID NO.119, SEQ ID NO. 121, SEQ ID NO. 123, SEQ ID NO. 125, SEQ ID NO. 127, SEQID NO. 129, SEQ ID NO. 130, SEQ ID NO. 131, SEQ ID NO. 132, SEQ ID NO.133, SEQ ID NO. 134, SEQ ID NO. 135, SEQ ID NO. 136, SEQ ID NO. 137, SEQID NO. 138, SEQ ID NO. 139, SEQ ID NO. 140, SEQ ID NO. 141, SEQ ID NO.142, SEQ ID NO. 143, and combinations thereof, and that has the lightchain variable domain sequence that is at least 95% identical, at least96% identical, at least 97% identical, at least 98% identical, or atleast 99% identical, to the amino acid sequence consisting of SEQ ID NO.2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO.12, SEQ ID NO. 14, SEQ ID NO. 16, SEQ ID NO. 18, SEQ ID NO. 20, SEQ IDNO. 22, SEQ ID NO. 24, SEQ ID NO. 26, SEQ ID NO. 28, SEQ ID NO. 30, SEQID NO. 32, SEQ ID NO. 34, SEQ ID NO. 36, SEQ ID NO. 38, SEQ ID NO. 40,SEQ ID NO. 42, SEQ ID NO. 44, SEQ ID NO. 46, SEQ ID NO. 48, SEQ ID NO.50, SEQ ID NO. 52, SEQ ID NO. 54, SEQ ID NO. 56, SEQ ID NO. 58, SEQ IDNO. 60, SEQ ID NO. 62, SEQ ID NO. 64, SEQ ID NO. 66, SEQ ID NO. 68, SEQID NO. 70, SEQ ID NO. 72, SEQ ID NO. 74, SEQ ID NO. 76, SEQ ID NO. 78,SEQ ID NO. 80, SEQ ID NO. 82, SEQ ID NO. 84, SEQ ID NO. 86, SEQ ID NO.88, SEQ ID NO. 90, SEQ ID NO. 92, SEQ ID NO. 94, SEQ ID NO. 96, SEQ IDNO. 98, SEQ ID NO. 100, SEQ ID NO. 102, SEQ ID NO. 104, SEQ ID NO. 106,SEQ ID NO. 108, SEQ ID NO. 110, SEQ ID NO. 112, SEQ ID NO. 114, SEQ IDNO. 116, SEQ ID NO. 118, SEQ ID NO. 120, SEQ ID NO. 122, SEQ ID NO. 124,SEQ ID NO. 126, SEQ ID NO. 128, and combinations thereof; and whereinthe single chain human antibody has the heavy chain variable domainsequence that is at least 95% identical, at least 96% identical, atleast 97% identical, at least 98% identical, or at least 99% identical,to the amino acid sequences selected from the group consisting of SEQ IDNO. 1, SEQ ID NO. 3, SEQ ID NO. 5, SEQ ID NO. 6, SEQ ID NO. 7, SEQ IDNO. 9, SEQ ID NO. 11, SEQ ID NO. 13, SEQ ID NO. 15, SEQ ID NO. 17, SEQID NO. 19, SEQ ID NO. 21, SEQ ID NO. 23, SEQ ID NO. 25, SEQ ID NO. 27,SEQ ID NO. 29, SEQ ID NO. 31, SEQ ID NO. 33, SEQ ID NO. 35, SEQ ID NO.37, SEQ ID NO. 39, SEQ ID NO. 41, SEQ ID NO. 43, SEQ ID NO. 45, SEQ IDNO. 47, SEQ ID NO. 49, SEQ ID NO. 51, SEQ ID NO. 53, SEQ ID NO. 55, SEQID NO. 57, SEQ ID NO. 59, SEQ ID NO. 61, SEQ ID NO. 63, SEQ ID NO. 65,SEQ ID NO. 67, SEQ ID NO. 69, SEQ ID NO. 71, SEQ ID NO. 73, SEQ ID NO.75, SEQ ID NO. 77, SEQ ID NO. 79, SEQ ID NO. 81, SEQ ID NO. 83, SEQ IDNO. 85, SEQ ID NO. 87, SEQ ID NO. 89, SEQ ID NO. 91, SEQ ID NO. 93, SEQID NO. 95, SEQ ID NO. 97, SEQ ID NO. 99, SEQ ID NO. 101, SEQ ID NO. 103,SEQ ID NO. 105, SEQ ID NO. 107, SEQ ID NO. 109, SEQ ID NO. 111, SEQ IDNO. 113, SEQ ID NO. 115, SEQ ID NO. 117, SEQ ID NO. 119, SEQ ID NO. 121,SEQ ID NO. 123, SEQ ID NO. 125, SEQ ID NO. 127, SEQ ID NO. 129, SEQ IDNO. 130, SEQ ID NO. 131, SEQ ID NO. 132, SEQ ID NO. 133, SEQ ID NO. 134,SEQ ID NO. 135, SEQ ID NO. 136, SEQ ID NO. 137, SEQ ID NO. 138, SEQ IDNO. 139, SEQ ID NO. 140, SEQ ID NO. 141, SEQ ID NO. 142, SEQ ID NO. 143,and combinations thereof, and that has the light chain variable domainsequence that is at least 95% identical, at least 96% identical, atleast 97% identical, at least 98% identical, or at least 99% identical,to the amino acid sequence consisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, SEQID NO. 16, SEQ ID NO. 18, SEQ ID NO. 20, SEQ ID NO. 22, SEQ ID NO. 24,SEQ ID NO. 26, SEQ ID NO. 28, SEQ ID NO. 30, SEQ ID NO. 32, SEQ ID NO.34, SEQ ID NO. 36, SEQ ID NO. 38, SEQ ID NO. 40, SEQ ID NO. 42, SEQ IDNO. 44, SEQ ID NO. 46, SEQ ID NO. 48, SEQ ID NO. 50, SEQ ID NO. 52, SEQID NO. 54, SEQ ID NO. 56, SEQ ID NO. 58, SEQ ID NO. 60, SEQ ID NO. 62,SEQ ID NO. 64, SEQ ID NO. 66, SEQ ID NO. 68, SEQ ID NO. 70, SEQ ID NO.72, SEQ ID NO. 74, SEQ ID NO. 76, SEQ ID NO. 78, SEQ ID NO. 80, SEQ IDNO. 82, SEQ ID NO. 84, SEQ ID NO. 86, SEQ ID NO. 88, SEQ ID NO. 90, SEQID NO. 92, SEQ ID NO. 94, SEQ ID NO. 96, SEQ ID NO. 98, SEQ ID NO. 100,SEQ ID NO. 102, SEQ ID NO. 104, SEQ ID NO. 106, SEQ ID NO. 108, SEQ IDNO. 110, SEQ ID NO. 112, SEQ ID NO. 114, SEQ ID NO. 116, SEQ ID NO. 118,SEQ ID NO. 120, SEQ ID NO. 122, SEQ ID NO. 124, SEQ ID NO. 126, SEQ IDNO. 128, and combinations thereof.

In one embodiment, the fully human antibody has both a heavy chain and alight chain wherein the antibody has a heavy chain/light chain variabledomain sequence selected from the group consisting of SEQ ID NO. 1/SEQID NO. 2 (called A1 herein), SEQ ID NO. 3/SEQ ID NO. 4 (called A4herein), SEQ ID NO. 5/SEQ ID NO. 6 (called A11 herein), SEQ ID NO. 7/SEQID NO. 8 (called B1 herein), SEQ ID NO. 9/SEQ ID NO. 10 (called B3herein), SEQ ID NO. 11/SEQ ID NO. 12 (called B12 herein), SEQ ID NO.13/SEQ ID NO. 14 (called C2 herein), SEQ ID NO. 15/SEQ ID NO. 16 (calledC3 herein), SEQ ID NO. 17/SEQ ID NO. 18 (called C7 herein), SEQ ID NO.19/SEQ ID NO. 20 (called C11 herein), SEQ ID NO. 21/SEQ ID NO. 22(called C12 herein), SEQ ID NO. 23/SEQ ID NO. 24 (called D1 herein), SEQID NO. 25/SEQ ID NO. 26 (called D4 herein), SEQ ID NO. 27/SEQ ID NO. 28(called D6 herein), SEQ ID NO. 29/SEQ ID NO. 30 (called D7 herein), SEQID NO. 31/SEQ ID NO. 32 (called D8 herein), SEQ ID NO. 33/SEQ ID NO. 34(called D10 herein), SEQ ID NO. 35/SEQ ID NO. 36 (called E2 herein), SEQID NO. 37/SEQ ID NO. 38 (called E5 herein), SEQ ID NO. 39/SEQ ID NO. 40(called E7 herein), SEQ ID NO. 41/SEQ ID NO. 42 (called F5 herein), SEQID NO. 43/SEQ ID NO. 44 (called F7 herein), SEQ ID NO. 45/SEQ ID NO. 46(called F11 herein), SEQ ID NO. 47/SEQ ID NO. 48 (called G1 herein), SEQID NO. 49/SEQ ID NO. 50 (called G2 herein), SEQ ID NO. 51/SEQ ID NO. 52(called G3 herein), SEQ ID NO. 53/SEQ ID NO. 54 (called G5 herein), SEQID NO. 55/SEQ ID NO. 56 (called G6 herein), SEQ ID NO. 57/SEQ ID NO. 58(called G8 herein), SEQ ID NO. 59/SEQ ID NO. 60 (called G12 herein), SEQID NO. 61/SEQ ID NO. 62 (called H4 herein), SEQ ID NO. 63/SEQ ID NO. 64(called H7 herein), SEQ ID NO. 65/SEQ ID NO. 66 (called H8 herein), SEQID NO. 67/SEQ ID NO. 68 (called H10 herein), SEQ ID NO. 69/SEQ ID NO. 70(called H11 herein), SEQ ID NO. 71/SEQ ID NO. 72 (called C3sh1A1herein), SEQ ID NO. 73/SEQ ID NO. 74 (called C3sh1A2 herein), SEQ ID NO.75/SEQ ID NO. 76 (called C3sh1A5 herein), SEQ ID NO. 77/SEQ ID NO. 78(called C3sh1A9 herein), SEQ ID NO. 79/SEQ ID NO. 80 (called C3sh1B2herein), SEQ ID NO. 81/SEQ ID NO. 82 (called C3sh1B4 herein), SEQ ID NO.83/SEQ ID NO. 84 (called C3sh1B6 herein), SEQ ID NO. 85/SEQ ID NO. 86(called C3sh1B9 herein), SEQ ID NO. 87/SEQ ID NO. 88 (called C3sh1C1herein), SEQ ID NO. 89/SEQ ID NO. 90 (called C3sh1C2 herein), SEQ ID NO.91/SEQ ID NO. 92 (called C3sh1C7 herein), SEQ ID NO. 93/SEQ ID NO. 94(called C3sh1D1 herein), SEQ ID NO. 95/SEQ ID NO. 96 (called C3sh1D4herein), SEQ ID NO. 97/SEQ ID NO. 98 (called C3sh1D6 herein), SEQ ID NO.99/SEQ ID NO. 100 (called C3sh1E2 herein), SEQ ID NO. 101/SEQ ID NO. 102(called C3sh1E7 herein), SEQ ID NO. 103/SEQ ID NO. 104 (called C3sh1E9herein), SEQ ID NO. 105/SEQ ID NO. 106 (called C3sh1F1 herein), SEQ IDNO. 107/SEQ ID NO. 108 (called C3sh1F10 herein), SEQ ID NO. 109/SEQ IDNO. 110 (called C3sh1F12 herein), SEQ ID NO. 111/SEQ ID NO. 112 (calledC3sh1F2 herein), SEQ ID NO. 113/SEQ ID NO. 114 (called C3sh1G1 herein),SEQ ID NO. 115/SEQ ID NO. 116 (called C3sh1G11 herein), SEQ ID NO.117/SEQ ID NO. 118 (called C3sh1G2 herein), SEQ ID NO. 119/SEQ ID NO.120 (called C3sh1G3 herein), SEQ ID NO. 121/SEQ ID NO. 122 (calledC3sh1G5 herein), SEQ ID NO. 123/SEQ ID NO. 124 (called C3sh1G8 herein),SEQ ID NO. 125/SEQ ID NO. 126 (called C3sh1H10 herein), SEQ ID NO.127/SEQ ID NO. 128 (called C3sh1H4 herein), SEQ ID NO. 129/SEQ ID NO. 28(called MA8 herein), SEQ ID NO. 130/SEQ ID NO. 28 (called MB1 herein),SEQ ID NO. 131/SEQ ID NO. 28 (called MB3 herein), SEQ ID NO. 132/SEQ IDNO. 28 (called MB10 herein), SEQ ID NO. 133/SEQ ID NO. 28 (called MB12herein), SEQ ID NO. 134/SEQ ID NO. 28 (called MC8 herein), SEQ ID NO.135/SEQ ID NO. 28 (called MD1 herein), SEQ ID NO. 136/SEQ ID NO. 28(called MD4 herein), SEQ ID NO. 137/SEQ ID NO. 28 (called MSA11 herein),SEQ ID NO. 138/SEQ ID NO. 28 (called MSB7 herein), SEQ ID NO. 139/SEQ IDNO. 28 (called MSD2 herein), SEQ ID NO. 140/SEQ ID NO. 28 (called MSE3herein), SEQ ID NO. 141/SEQ ID NO. 28 (called MSE5 herein), SEQ ID NO.142/SEQ ID NO. 28 (called MSC8 herein), SEQ ID NO. 143/SEQ ID NO. 28(called MSH1 herein), and combinations thereof. Preferably, the fullyhuman single chain antibody has both a heavy chain variable domainregion and a light chain variable domain region, wherein the singlechain fully human antibody has a heavy chain/light chain variable domainsequence selected from the group consisting of SEQ ID NO. 1/SEQ ID NO.2, SEQ ID NO. 3/SEQ ID NO. 4, SEQ ID NO. 5/SEQ ID NO. 6, SEQ ID NO.7/SEQ ID NO. 8, SEQ ID NO. 9/SEQ ID NO. 10, SEQ ID NO. 11/SEQ ID NO. 12,SEQ ID NO. 13/SEQ ID NO. 14, SEQ ID NO. 15/SEQ ID NO. 16, SEQ ID NO.17/SEQ ID NO. 18, SEQ ID NO. 19/SEQ ID NO. 20, SEQ ID NO. 21/SEQ ID NO.22, SEQ ID NO. 23/SEQ ID NO. 24, SEQ ID NO. 25/SEQ ID NO. 26, SEQ ID NO.27/SEQ ID NO. 28, SEQ ID NO. 29/SEQ ID NO. 30, SEQ ID NO. 31/SEQ ID NO.32, SEQ ID NO. 33/SEQ ID NO. 34, SEQ ID NO. 35/SEQ ID NO. 36, SEQ ID NO.37/SEQ ID NO. 38, SEQ ID NO. 39/SEQ ID NO. 40, SEQ ID NO. 41/SEQ ID NO.42, and combinations thereof.

Preferably, the disease is selected from the group consisting ofcancers, autoimmune diseases and viral infections.

In certain embodiments, the anti-CD137 antibody, or antigen-bindingfragment thereof, of the invention has a K_(D) of at least 1×10⁻⁶M. Inother embodiments, the anti-CD137 antibody, or antigen-binding fragmentthereof, of the invention has a K_(D) of at least 1×10⁻⁷M. In otherembodiments, the anti-CD137 antibody, or antigen-binding fragmentthereof, of the invention has a K_(D) of at least 1×10⁻⁸M.

In certain embodiments, the anti-CD137 antibody is an IgG1 isotype. Inother embodiments, the anti-CD137 antibody is an IgG4 isotype.

In certain embodiments, the anti-CD137 antibody, or antigen-bindingfragment, described herein is recombinant.

The invention also provides pharmaceutical compositions comprising aneffective amount of an anti-CD137 antibodies or fragments disclosedherein, and a pharmaceutically acceptable carrier.

In certain embodiments, the invention features a method of treatingcancer in a human subject in need thereof, comprising administering aneffective amount of an anti-CD137 antibody, or antigen-binding fragmentthereof, disclosed herein to the subject, such that cancer is treated.Examples of cancer that may be treated include, but are not limited to,ovarian cancer, colorectal cancer, melanoma, hepatocellular carcinoma,renal cancer, breast cancer, head and neck cancer, lung cancer and livercancer.

DESCRIPTION OF THE DRAWINGS

FIG. 1A is a graph that shows functional activity of the listedanti-CD137 antibodies by their ability to augment T cell activation. Tomeasure cell activation, the cells were labeled with FITC anti-humanCD25 after three days of culture. The percentage of cells positive forCD25 expression was measured by flow cytometry. The level of CD25expression was higher in the cultures where anti-CD137 antibodies hadbeen added. cIg is a control immunoglobulin which is not specific forCD137.

FIG. 1B is a graph that shows the results of FIG. 1A, normalizedrelative to the cultures receiving no antibody. A notable level ofaugmentation was detected in cultures which received the testedanti-CD137 antibodies, and in particular the D6 and C7 antibodies.

FIG. 2 graphically depicts cross-reactivity studies determining whetheranti-CD137 antibodies D6, MB3, MSC8, and MB12 are able to bind humanand/or murine CD137. The results show that each of these antibodies isspecific for human CD137, as none of the antibodies showedcross-reactivity to murine CD137.

FIG. 3A and FIG. 3B graphically depict results from an in vitroexperiment determining cell activation. The percentage of cells positivefor CD25 expression was measured by flow cytometry. The level of CD25expression was higher in the cultures where anti-CD137 antibodies hadbeen added. FIG. 3B provides the normalized results of FIG. 3A. cIg is acontrol immunoglobulin which is not specific for CD137.

DETAILED DESCRIPTION Definitions

The terms “peptide,” “polypeptide” and “protein” each refers to amolecule comprising two or more amino acid residues joined to each otherby peptide bonds. These terms encompass, e.g., native and artificialproteins, protein fragments and polypeptide analogs (such as muteins,variants, and fusion proteins) of a protein sequence as well aspost-translationally, or otherwise covalently or non-covalently,modified proteins. A peptide, polypeptide, or protein may be monomericor polymeric.

A “variant” of a polypeptide (for example, a variant of an antibody)comprises an amino acid sequence wherein one or more amino acid residuesare inserted into, deleted from and/or substituted into the amino acidsequence relative to another polypeptide sequence. Disclosed variantsinclude, for example, fusion proteins.

A “derivative” of a polypeptide is a polypeptide (e.g., an antibody)that has been chemically modified, e.g., via conjugation to anotherchemical moiety (such as, for example, polyethylene glycol or albumin,e.g., human serum albumin), phosphorylation, and glycosylation. Unlessotherwise indicated, the term “antibody” includes, in addition toantibodies comprising two full-length heavy chains and two full-lengthlight chains, derivatives, variants, fragments, and muteins thereof,examples of which are described below.

An “antigen binding protein” is a protein comprising a portion thatbinds to an antigen and, optionally, a scaffold or framework portionthat allows the antigen binding portion to adopt a confirmation thatpromotes binding of the antigen binding protein to the antigen. Examplesof antigen binding proteins include antibodies, antibody fragments(e.g., an antigen binding portion of an antibody), antibody derivatives,and antibody analogs. The antigen binding protein can comprise, forexample, an alternative protein scaffold or artificial scaffold withgrafted CDRs or CDR derivatives. Such scaffolds include, but are notlimited to, antibody-derived scaffolds comprising mutations introducedto, for example, stabilize the three-dimensional structure of theantigen binding protein as well as wholly synthetic scaffoldscomprising, for example, a biocompatible polymer. See, for example,Korndorfer et al., 2003, Proteins: Structure, Function, andBioinformatics, Volume 53, Issue 1:121-129; Roque et al., 2004,Biotechnol. Prog. 20:639-654. In addition, peptide antibody mimetics(“PAMs”) can be used, as well as scaffolds based on antibody mimeticsutilizing fibronection components as a scaffold.

An antigen binding protein can have, for example, the structure of animmunoglobulin. An “immunoglobulin” is a tetrameric molecule composed oftwo identical pairs of polypeptide chains, each pair having one “light”(about 25 kDa) and one “heavy” chain (about 50-70 kDa). Theamino-terminal portion of each chain includes a variable region of about100 to 110 or more amino acids primarily responsible for antigenrecognition. The carboxy-terminal portion of each chain defines aconstant region primarily responsible for effector function. Human lightchains are classified as kappa or lambda light chains. Heavy chains areclassified as mu, delta, gamma, alpha, or epsilon, and define theantibody's isotype as IgM, IgD, IgG, IgA, and IgE, respectively.Preferably, the anti-EGFR antibodies disclosed herein are characterizedby their variable domain region sequences in the heavy V_(H) and lightV_(L) amino acid sequences. The preferred antibody is A6 which is akappa IgG antibody. Within light and heavy chains, the variable andconstant regions are joined by a “J” region of about 12 or more aminoacids, with the heavy chain also including a “D” region of about 10 moreamino acids. See generally, Fundamental Immunology Ch. 7 (Paul, W., ed.,2nd ed. Raven Press, N.Y. (1989)). The variable regions of eachlight/heavy chain pair form the antibody binding site such that anintact immunoglobulin has two binding sites.

The variable regions of immunoglobulin chains exhibit the same generalstructure of relatively conserved framework regions (FR) joined by threehypervariable regions, also called complementarity determining regionsor CDRs. From N-terminus to C-terminus, both light and heavy chainscomprise the domains FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. Theassignment of amino acids to each domain is in accordance with thedefinitions of Kabat et al. in Sequences of Proteins of ImmunologicalInterest, 5th Ed., US Dept. of Health and Human Services, PHS, NIH, NIHPublication no. 91-3242, 1991. Other numbering systems for the aminoacids in immunoglobulin chains include IMGT™ (internationalImMunoGeneTics information system; Lefranc et al, Dev. Comp. Immunol.29:185-203; 2005) and AHo (Honegger and Pluckthun, J. Mol. Biol.309(3):657-670; 2001).

An “antibody” refers to an intact immunoglobulin or to an antigenbinding portion thereof that competes with the intact antibody forspecific binding, unless otherwise specified. In one embodiment, anantibody comprises a heavy chain variable domain, a light chain variabledomain, a light chain constant region (C_(L)), and heavy chain constantregions C_(H1), C_(H2) and C_(H3). The heavy and light chain variabledomain sequences may be selected from those described herein in SEQ IDNos: 1 to 143.

Antigen binding portions of an antibody may be produced by recombinantDNA techniques or by enzymatic or chemical cleavage of intactantibodies. Antigen binding portions include, inter alia, Fab, Fab′,F(ab′)2, Fv, domain antibodies (dAbs), and complementarity determiningregion (CDR) fragments, single-chain antibodies (scFv), chimericantibodies, diabodies, triabodies, tetrabodies, and polypeptides thatcontain at least a portion of an immunoglobulin that is sufficient toconfer specific antigen binding to the polypeptide.

In certain embodiments, antibodies can be obtained from sources such asserum or plasma that contain immunoglobulins having varied antigenicspecificity. If such antibodies are subjected to affinity purification,they can be enriched for a particular antigenic specificity. Suchenriched preparations of antibodies usually are made of less than about10% antibody having specific binding activity for the particularantigen. Subjecting these preparations to several rounds of affinitypurification can increase the proportion of antibody having specificbinding activity for the antigen. Antibodies prepared in this manner areoften referred to as “monospecific.”

The term “monospecific”, as used herein, refers to an antibody thatdisplays an affinity for one particular epitope. Monospecific antibodypreparations can be made up of about 10%, 20%, 30%, 40%, 50%, 60%, 70%,75%, 80%, 85%, 90%, 95%, 97%, 99%, or 99.9% antibody having specificbinding activity for the particular antigen.

An “antibody fragment” or “antigen binding fragment of an antibody”comprises a portion of an intact antibody, and preferably comprises theantibody antigen binding or variable domains. Examples of an antibodyfragment include a Fab, an Fab′, an F(ab′)2, an Fv fragment, and alinear antibody.

A Fab fragment is a monovalent fragment having the V_(L), V_(H), C_(L)and C_(H1) domains; a F(ab′)₂ fragment is a bivalent fragment having twoFab fragments linked by a disulfide bridge at the hinge region; a Fdfragment has the V_(H) and C_(H1) domains; an Fv fragment has the V_(L)and V_(H) domains of a single arm of an antibody; and a dAb fragment hasa V_(H) domain, a V_(L) domain, or an antigen-binding fragment of aV_(H) or V_(L) domain (U.S. Pat. Nos. 6,846,634; 6,696,245, US App. Pub.20/0202512; 2004/0202995; 2004/0038291; 2004/0009507;2003/0039958, andWard et al., Nature 341:544-546, 1989).

A single-chain antibody (scFv) is an antibody fragment in which a V_(L)and a V_(H) region are joined via a linker (e.g., a synthetic sequenceof amino acid residues) to form a continuous protein chain wherein thelinker is long enough to allow the protein chain to fold back on itselfand form a monovalent antigen binding site (see, e.g., Bird et al.,1988, Science 242:423-26 and Huston et al., 1988, Proc. Natl. Acad. Sci.USA 85:5879-83).

Diabodies are bivalent antibodies comprising two polypeptide chains,wherein each polypeptide chain comprises V_(H) and V_(L) domains joinedby a linker that is too short to allow for pairing between two domainson the same chain, thus allowing each domain to pair with acomplementary domain on another polypeptide chain (see, e.g., Holligeret al., 1993, Proc. Natl. Acad. Sci. USA 90:6444-48, and Poljak et al.,1994, Structure 2:1121-23). If the two polypeptide chains of a diabodyare identical, then a diabody resulting from their pairing will have twoidentical antigen binding sites. Polypeptide chains having differentsequences can be used to make a diabody with two different antigenbinding sites. Similarly, tribodies and tetrabodies are antibodiescomprising three and four polypeptide chains, respectively, and formingthree and four antigen binding sites, respectively, which can be thesame or different.

An antigen binding protein, such as an antibody, may have one or morebinding sites. If there is more than one binding site, the binding sitesmay be identical to one another or may be different. For example, anaturally occurring human immunoglobulin typically has two identicalbinding sites, while a “bispecific” or “bifunctional” antibody has twodifferent binding sites.

The term “human antibody” includes all antibodies that have one or morevariable and constant regions derived from human immunoglobulinsequences. In one embodiment, all of the variable and constant domainsof the antibody are derived from human immunoglobulin sequences(referred to as a “fully human antibody”). These antibodies may beprepared in a variety of ways, examples of which are described below,including through the immunization with an antigen of interest of amouse that is genetically modified to express antibodies derived fromhuman heavy and/or light chain-encoding genes. In a preferredembodiment, a fully human antibody is made using recombinant methodssuch that the glycosylation pattern of the antibody is different than anantibody having the same sequence if it were to exist in nature.

A “humanized antibody” has a sequence that differs from the sequence ofan antibody derived from a non-human species by one or more amino acidsubstitutions, deletions, and/or additions, such that the humanizedantibody is less likely to induce an immune response, and/or induces aless severe immune response, as compared to the non-human speciesantibody, when it is administered to a human subject. In one embodiment,certain amino acids in the framework and constant domains of the heavyand/or light chains of the non-human species antibody are mutated toproduce the humanized antibody. In another embodiment, the constantdomain(s) from a human antibody are fused to the variable domain(s) of anon-human species. In another embodiment, one or more amino acidresidues in one or more CDR sequences of a non-human antibody arechanged to reduce the likely immunogenicity of the non-human antibodywhen it is administered to a human subject, wherein the changed aminoacid residues either are not critical for immunospecific binding of theantibody to its antigen, or the changes to the amino acid sequence thatare made are conservative changes, such that the binding of thehumanized antibody to the antigen is not significantly worse than thebinding of the non-human antibody to the antigen. Examples of how tomake humanized antibodies may be found in U.S. Pat. Nos. 6,054,297,5,886,152 and 5,877,293.

The term “chimeric antibody” refers to an antibody that contains one ormore regions from one antibody and one or more regions from one or moreother antibodies. In one embodiment, one or more of the CDRs are derivedfrom a human anti-CD137 antibody. In another embodiment, all of the CDRsare derived from a human anti-CD137 antibody. In another embodiment, theCDRs from more than one human anti-CD137 antibodies are mixed andmatched in a chimeric antibody. For instance, a chimeric antibody maycomprise a CDR1 from the light chain of a first human anti-PAR-2antibody, a CDR2 and a CDR3 from the light chain of a second humananti-CD137 antibody, and the CDRs from the heavy chain from a thirdanti-CD137 antibody. Other combinations are possible.

Further, the framework regions may be derived from one of the sameanti-CD137 antibodies, from one or more different antibodies, such as ahuman antibody, or from a humanized antibody. In one example of achimeric antibody, a portion of the heavy and/or light chain isidentical with, homologous to, or derived from an antibody from aparticular species or belonging to a particular antibody class orsubclass, while the remainder of the chain(s) is/are identical with,homologous to, or derived from an antibody (-ies) from another speciesor belonging to another antibody class or subclass. Also included arefragments of such antibodies that exhibit the desired biologicalactivity (i.e., the ability to specifically bind CD137).

An “agonist antibody” as used herein, is an antibody that induces orincreases the biological activity of an antigen (for example, CD137) towhich the antibody binds. An agonist may, for example, facilitate areceptor's phosphorylation due to binding of the receptor to a ligand ormay activate or grow cells activated by the receptor. In one embodiment,the antibodies of the invention are agonist anti-CD137 antibodies.

A “CDR grafted antibody” is an antibody comprising one or more CDRsderived from an antibody of a particular species or isotype and theframework of another antibody of the same or different species orisotype.

A “multi-specific antibody” is an antibody that recognizes more than oneepitope on one or more antigens. A subclass of this type of antibody isa “bi-specific antibody” which recognizes two distinct epitopes on thesame or different antigens.

An antigen binding protein “specifically binds” to an antigen (e.g.,human CD137) if it binds to the antigen with a dissociation constant of1 nanomolar or less.

An “antigen binding domain,” “antigen binding region,” or “antigenbinding site” is a portion of an antigen binding protein that containsamino acid residues (or other moieties) that interact with an antigenand contribute to the antigen binding protein's specificity and affinityfor the antigen. For an antibody that specifically binds to its antigen,this will include at least part of at least one of its CDR domains.

The term “Fc polypeptide” includes native and mutein forms ofpolypeptides derived from the Fc region of an antibody. Truncated formsof such polypeptides containing the hinge region that promotesdimerization also are included. Fusion proteins comprising Fc moieties(and oligomers formed therefrom) offer the advantage of facilepurification by affinity chromatography over Protein A or Protein Gcolumns.

An “epitope” is the portion of a molecule that is bound by an antigenbinding protein (e.g., by an antibody). An epitope can comprisenon-contiguous portions of the molecule (e.g., in a polypeptide, aminoacid residues that are not contiguous in the polypeptide's primarysequence but that, in the context of the polypeptide's tertiary andquaternary structure, are near enough to each other to be bound by anantigen binding protein).

The “percent identity” or “percent homology” of two polynucleotide ortwo polypeptide sequences is determined by comparing the sequences usingthe GAP computer program (a part of the GCG Wisconsin Package, version10.3 (Accelrys, San Diego, Calif.)) using its default parameters.

The terms “polynucleotide,” “oligonucleotide” and “nucleic acid” areused interchangeably throughout and include DNA molecules (e.g., cDNA orgenomic DNA), RNA molecules (e.g., mRNA), analogs of the DNA or RNAgenerated using nucleotide analogs (e.g., peptide nucleic acids andnon-naturally occurring nucleotide analogs), and hybrids thereof. Thenucleic acid molecule can be single-stranded or double-stranded. In oneembodiment, the nucleic acid molecules of the invention comprise acontiguous open reading frame encoding an antibody, or a fragment,derivative, mutein, or variant thereof.

Two single-stranded polynucleotides are “the complement” of each otherif their sequences can be aligned in an anti-parallel orientation suchthat every nucleotide in one polynucleotide is opposite itscomplementary nucleotide in the other polynucleotide, without theintroduction of gaps, and without unpaired nucleotides at the 5′ or the3′ end of either sequence. A polynucleotide is “complementary” toanother polynucleotide if the two polynucleotides can hybridize to oneanother under moderately stringent conditions. Thus, a polynucleotidecan be complementary to another polynucleotide without being itscomplement.

A “vector” is a nucleic acid that can be used to introduce anothernucleic acid linked to it into a cell. One type of vector is a“plasmid,” which refers to a linear or circular double stranded DNAmolecule into which additional nucleic acid segments can be ligated.Another type of vector is a viral vector (e.g., replication defectiveretroviruses, adenoviruses and adeno-associated viruses), whereinadditional DNA segments can be introduced into the viral genome. Certainvectors are capable of autonomous replication in a host cell into whichthey are introduced (e.g., bacterial vectors comprising a bacterialorigin of replication and episomal mammalian vectors). Other vectors(e.g., non-episomal mammalian vectors) are integrated into the genome ofa host cell upon introduction into the host cell, and thereby arereplicated along with the host genome. An “expression vector” is a typeof vector that can direct the expression of a chosen polynucleotide.

A nucleotide sequence is “operably linked” to a regulatory sequence ifthe regulatory sequence affects the expression (e.g., the level, timing,or location of expression) of the nucleotide sequence. A “regulatorysequence” is a nucleic acid that affects the expression (e.g., thelevel, timing, or location of expression) of a nucleic acid to which itis operably linked. The regulatory sequence can, for example, exert itseffects directly on the regulated nucleic acid, or through the action ofone or more other molecules (e.g., polypeptides that bind to theregulatory sequence and/or the nucleic acid). Examples of regulatorysequences include promoters, enhancers and other expression controlelements (e.g., polyadenylation signals). Further examples of regulatorysequences are described in, for example, Goeddel, 1990, Gene ExpressionTechnology: Methods in Enzymology 185, Academic Press, San Diego, Calif.and Baron et al., 1995, Nucleic Acids Res. 23:3605-06.

A “host cell” is a cell that can be used to express a nucleic acid,e.g., a nucleic acid of the invention. A host cell can be a prokaryote,for example, E. coli, or it can be a eukaryote, for example, asingle-celled eukaryote (e.g., a yeast or other fungus), a plant cell(e.g., a tobacco or tomato plant cell), an animal cell (e.g., a humancell, a monkey cell, a hamster cell, a rat cell, a mouse cell, or aninsect cell) or a hybridoma. Examples of host cells include the COS-7line of monkey kidney cells (ATCC CRL 1651) (see Gluzman et al., 1981,Cell 23:175), L cells, C127 cells, 3T3 cells (ATCC CCL 163), Chinesehamster ovary (CHO) cells or their derivatives such as Veggie CHO andrelated cell lines which grow in serum-free media (see Rasmussen et al.,1998, Cytotechnology 28:31) or CHO strain DX-B11, which is deficient inDHFR (see Urlaub et al., 1980, Proc. Natl. Acad. Sci. USA 77:4216-20),HeLa cells, BHK (ATCC CRL 10) cell lines, the CV1/EBNA cell line derivedfrom the African green monkey kidney cell line CV1 (ATCC CCL 70) (seeMcMahan et al., 1991, EMBO J. 10:2821), human embryonic kidney cellssuch as 293,293 EBNA or MSR 293, human epidermal A431 cells, humanColo205 cells, other transformed primate cell lines, normal diploidcells, cell strains derived from in vitro culture of primary tissue,primary explants, HL-60, U937, HaK or Jurkat cells. In one embodiment, ahost cell is a mammalian host cell, but is not a human host cell.Typically, a host cell is a cultured cell that can be transformed ortransfected with a polypeptide-encoding nucleic acid, which can then beexpressed in the host cell. The phrase “recombinant host cell” can beused to denote a host cell that has been transformed or transfected witha nucleic acid to be expressed. A host cell also can be a cell thatcomprises the nucleic acid but does not express it at a desired levelunless a regulatory sequence is introduced into the host cell such thatit becomes operably linked with the nucleic acid. It is understood thatthe term host cell refers not only to the particular subject cell butalso to the progeny or potential progeny of such a cell. Because certainmodifications may occur in succeeding generations due to, e.g., mutationor environmental influence, such progeny may not, in fact, be identicalto the parent cell, but are still included within the scope of the termas used herein.

The term “recombinant antibody” refers to an antibody that is expressedfrom a cell or cell line transfected with an expression vector (orpossibly more than one expression vector, e.g., two expression vectors)comprising at least the coding sequence of the antibody, where saidcoding sequence is not naturally associated with the cell. In oneembodiment, a recombinant antibody has a glycosylation pattern that isdifferent than the glycosylation pattern of an antibody having the samesequence if it were to exist in nature. In one embodiment, a recombinantantibody is expressed in a mammalian host cell which is not a human hostcell. Notably, individual mammalian host cells have unique glycosylationpatterns.

The term “effective amount” as used herein, refers to that amount of anantibody, or an antigen binding portion thereof, that binds CD137, whichis sufficient to effect treatment, prognosis or diagnosis of a diseaseassociated with CD137 dependent signaling, as described herein, whenadministered to a subject. Therapeutically effective amounts ofantibodies provided herein, when used alone or in combination, will varydepending upon the relative activity of the antibodies and combinations(e.g., in inhibiting cell growth) and depending upon the subject anddisease condition being treated, the weight and age of the subject, theseverity of the disease condition, the manner of administration and thelike, which can readily be determined by one of ordinary skill in theart.

The term “isolated” refers to a protein (e.g., an antibody) that issubstantially free of other cellular material and/or chemicals. In oneembodiment, an isolated antibody is substantially free of other proteinsfrom the same species. In one embodiment, an isolated antibody isexpressed by a cell from a different species and is substantially freeof other proteins from the different species. A protein may be renderedsubstantially free of naturally associated components (or componentsassociated with the cellular expression system used to produce theantibody) by isolation, using protein purification techniques well knownin the art. In one embodiment, the antibodies, or antigen bindingfragments, of the invention are isolated.

CD137 Antigen Binding Proteins

The present invention pertains to CD137 binding proteins, particularlyanti-CD137 antibodies, or antigen-binding portions thereof, that bindCD137, and uses thereof. Various aspects of the invention relate toantibodies and antibody fragments, pharmaceutical compositions, nucleicacids, recombinant expression vectors, and host cells for making suchantibodies and fragments. Methods of using the antibodies of theinvention to detect human CD137, to stimulate CD137 activity, either invitro or in vivo, and to prevent or treat disorders such as cancer arealso encompassed by the invention.

As described in Table 5 below, included in the invention are novelantibody heavy and light chain variable regions that are specific toCD137. In one embodiment, the invention provides an anti-CD137 antibody,or an antigen-binding fragment thereof, that comprises a heavy chainhaving a variable domain comprising an amino acid sequence as set forthin any one of SEQ ID Nos: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25,27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61,63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97,99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125,127, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141,142 and 143. In one embodiment, the invention provides an anti-CD137antibody, or an antigen-binding fragment thereof, that comprises a lightchain having a variable domain comprising an amino acid sequence as setforth in any one of SEQ ID Nos: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22,24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58,60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94,96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124,126 and 128. In one embodiment, the invention provides an anti-CD137antibody, or an antigen-binding fragment thereof, that comprises a lightchain having a variable domain comprising an amino acid sequence as setforth in any one of SEQ ID Nos: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22,24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58,60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94,96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124,126 and 128; and a heavy chain having a variable domain comprising anamino acid sequence as set forth in any one of SEQ ID Nos: 1, 3, 5, 7,9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43,45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79,81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111,113, 115, 117, 119, 121, 123, 125, 127, 129, 130, 131, 132, 133, 134,135, 136, 137, 138, 139, 140, 141, 142 and 143.

Complementarity determining regions (CDRs) are known as hypervariableregions both in the light chain and the heavy chain variable domains.The more highly conserved portions of variable domains are called theframework (FR). Complementarity determining regions (CDRs) and frameworkregions (FR) of a given antibody may be identified using the systemdescribed by Kabat et al. supra; Lefranc et al., supra and/or Honeggerand Pluckthun, supra. Also familiar to those in the art is the numberingsystem described in Kabat et al. (1991, NIH Publication 91-3242,National Technical Information Service, Springfield, Va.). In thisregard Kabat et al. defined a numbering system for variable domainsequences that is applicable to any antibody. One of ordinary skill inthe art can unambiguously assign this system of “Kabat numbering” to anyvariable domain amino acid sequence, without reliance on anyexperimental data beyond the sequence itself.

In certain embodiments, the present invention provides an anti-CD137antibody comprising the CDRs of the heavy and light chain variabledomains described in Table 5 (SEQ ID Nos: 1 to 143). For example, theinvention provides an anti-CD137 antibody, or antigen-binding fragmentthereof, comprising a heavy chain variable region having the CDRsdescribed in an amino acid sequence as set forth in any one of SEQ IDNos: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35,37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71,73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105,107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 130, 131,132, 133, 134, 135, 136, 137, 139, 140, 141, 142 and 143. In oneembodiment, the invention provides an anti-CD137 antibody, orantigen-binding fragment thereof, comprising a light chain variableregion having the CDRs described in an amino acid sequence as set forthin any one of SEQ ID Nos:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26,28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62,64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98,100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126 and128. In one embodiment, the invention provides an anti-CD137 antibody,or antigen-binding fragment thereof, comprising a light chain variableregion having the CDRs described in an amino acid sequence as set forthin any one of SEQ ID Nos: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24,26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60,62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96,98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126and 128; and a heavy chain variable region having the CDRs described inan amino acid sequence as set forth in any one of SEQ ID Nos: 1, 3, 5,7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41,43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77,79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109,111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 130, 131, 132, 133,134, 135, 136, 137, 139, 140, 141, 142 and 143.

One or more CDRs may be incorporated into a molecule either covalentlyor noncovalently to make it an antigen binding protein.

An antigen binding protein may incorporate the CDR(s) as part of alarger polypeptide chain, may covalently link the CDR(s) to anotherpolypeptide chain, or may incorporate the CDR(s) noncovalently. The CDRspermit the antigen binding protein to specifically bind to a particularantigen of interest.

In one embodiment, the present disclosure provides a fully humanantibody of an IgG class that binds to a CD137 epitope with a bindingaffinity of 10⁻⁶M or less, that has a heavy chain variable domainsequence that is at least 95% identical to the amino acid sequencesselected from the group consisting of SEQ ID NO. 1, SEQ ID NO. 3, SEQ IDNO. 5, SEQ ID NO. 6, SEQ ID NO. 7, SEQ ID NO. 9, SEQ ID NO. 11, SEQ IDNO. 13, SEQ ID NO. 15, SEQ ID NO. 17, SEQ ID NO. 19, SEQ ID NO. 21, SEQID NO. 23, SEQ ID NO. 25, SEQ ID NO. 27, SEQ ID NO. 29, SEQ ID NO. 31,SEQ ID NO. 33, SEQ ID NO. 35, SEQ ID NO. 37, SEQ ID NO. 39, SEQ ID NO.41, SEQ ID NO. 43, SEQ ID NO. 45, SEQ ID NO. 47, SEQ ID NO. 49, SEQ IDNO. 51, SEQ ID NO. 53, SEQ ID NO. 55, SEQ ID NO. 57, SEQ ID NO. 59, SEQID NO. 61, SEQ ID NO. 63, SEQ ID NO. 65, SEQ ID NO. 67, SEQ ID NO. 69,SEQ ID NO. 71, SEQ ID NO. 73, SEQ ID NO. 75, SEQ ID NO. 77, SEQ ID NO.79, SEQ ID NO. 81, SEQ ID NO. 83, SEQ ID NO. 85, SEQ ID NO. 87, SEQ IDNO. 89, SEQ ID NO. 91, SEQ ID NO. 93, SEQ ID NO. 95, SEQ ID NO. 97, SEQID NO. 99, SEQ ID NO. 101, SEQ ID NO. 103, SEQ ID NO. 105, SEQ ID NO.107, SEQ ID NO. 109, SEQ ID NO. 111, SEQ ID NO. 113, SEQ ID NO. 115, SEQID NO. 117, SEQ ID NO. 119, SEQ ID NO. 121, SEQ ID NO. 123, SEQ ID NO.125, SEQ ID NO. 127, SEQ ID NO. 129, SEQ ID NO. 130, SEQ ID NO. 131, SEQID NO. 132, SEQ ID NO. 133, SEQ ID NO. 134, SEQ ID NO. 135, SEQ ID NO.136, SEQ ID NO. 137, SEQ ID NO. 138, SEQ ID NO. 139, SEQ ID NO. 140, SEQID NO. 141, SEQ ID NO. 142, SEQ ID NO. 143, and combinations thereof,and that has a light chain variable domain sequence that is at least 95%identical to the amino acid sequence consisting of SEQ ID NO. 2, SEQ IDNO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ IDNO. 14, SEQ ID NO. 16, SEQ ID NO. 18, SEQ ID NO. 20, SEQ ID NO. 22, SEQID NO. 24, SEQ ID NO. 26, SEQ ID NO. 28, SEQ ID NO. 30, SEQ ID NO. 32,SEQ ID NO. 34, SEQ ID NO. 36, SEQ ID NO. 38, SEQ ID NO. 40, SEQ ID NO.42, SEQ ID NO. 44, SEQ ID NO. 46, SEQ ID NO. 48, SEQ ID NO. 50, SEQ IDNO. 52, SEQ ID NO. 54, SEQ ID NO. 56, SEQ ID NO. 58, SEQ ID NO. 60, SEQID NO. 62, SEQ ID NO. 64, SEQ ID NO. 66, SEQ ID NO. 68, SEQ ID NO. 70,SEQ ID NO. 72, SEQ ID NO. 74, SEQ ID NO. 76, SEQ ID NO. 78, SEQ ID NO.80, SEQ ID NO. 82, SEQ ID NO. 84, SEQ ID NO. 86, SEQ ID NO. 88, SEQ IDNO. 90, SEQ ID NO. 92, SEQ ID NO. 94, SEQ ID NO. 96, SEQ ID NO. 98, SEQID NO. 100, SEQ ID NO. 102, SEQ ID NO. 104, SEQ ID NO. 106, SEQ ID NO.108, SEQ ID NO. 110, SEQ ID NO. 112, SEQ ID NO. 114, SEQ ID NO. 116, SEQID NO. 118, SEQ ID NO. 120, SEQ ID NO. 122, SEQ ID NO. 124, SEQ ID NO.126, SEQ ID NO. 128, and combinations thereof.

In one embodiment, the fully human antibody has both a heavy chain and alight chain wherein the antibody has a heavy chain/light chain variabledomain sequence selected from the group consisting of SEQ ID NO. 1/SEQID NO. 2 (called A1 herein), SEQ ID NO. 3/SEQ ID NO. 4 (called A4herein), SEQ ID NO. 5/SEQ ID NO. 6 (called A11 herein), SEQ ID NO. 7/SEQID NO. 8 (called B1 herein), SEQ ID NO. 9/SEQ ID NO. 10 (called B3herein), SEQ ID NO. 11/SEQ ID NO. 12 (called B12 herein), SEQ ID NO.13/SEQ ID NO. 14 (called C2 herein), SEQ ID NO. 15/SEQ ID NO. 16 (calledC3 herein), SEQ ID NO. 17/SEQ ID NO. 18 (called C7 herein), SEQ ID NO.19/SEQ ID NO. 20 (called C11 herein), SEQ ID NO. 21/SEQ ID NO. 22(called C12 herein), SEQ ID NO. 23/SEQ ID NO. 24 (called D1 herein), SEQID NO. 25/SEQ ID NO. 26 (called D4 herein), SEQ ID NO. 27/SEQ ID NO. 28(called D6 herein), SEQ ID NO. 29/SEQ ID NO. 30 (called D7 herein), SEQID NO. 31/SEQ ID NO. 32 (called D8 herein), SEQ ID NO. 33/SEQ ID NO. 34(called D10 herein), SEQ ID NO. 35/SEQ ID NO. 36 (called E2 herein), SEQID NO. 37/SEQ ID NO. 38 (called E5 herein), SEQ ID NO. 39/SEQ ID NO. 40(called E7 herein), SEQ ID NO. 41/SEQ ID NO. 42 (called F5 herein), SEQID NO. 43/SEQ ID NO. 44 (called F7 herein), SEQ ID NO. 45/SEQ ID NO. 46(called F11 herein), SEQ ID NO. 47/SEQ ID NO. 48 (called G1 herein), SEQID NO. 49/SEQ ID NO. 50 (called G2 herein), SEQ ID NO. 51/SEQ ID NO. 52(called G3 herein), SEQ ID NO. 53/SEQ ID NO. 54 (called G5 herein), SEQID NO. 55/SEQ ID NO. 56 (called G6 herein), SEQ ID NO. 57/SEQ ID NO. 58(called G8 herein), SEQ ID NO. 59/SEQ ID NO. 60 (called G12 herein), SEQID NO. 61/SEQ ID NO. 62 (called H4 herein), SEQ ID NO. 63/SEQ ID NO. 64(called H7 herein), SEQ ID NO. 65/SEQ ID NO. 66 (called H8 herein), SEQID NO. 67/SEQ ID NO. 68 (called H10 herein), SEQ ID NO. 69/SEQ ID NO. 70(called H11 herein), SEQ ID NO. 71/SEQ ID NO. 72 (called C3sh1A1herein), SEQ ID NO. 73/SEQ ID NO. 74 (called C3sh1A2 herein), SEQ ID NO.75/SEQ ID NO. 76 (called C3sh1A5 herein), SEQ ID NO. 77/SEQ ID NO. 78(called C3sh1A9 herein), SEQ ID NO. 79/SEQ ID NO. 80 (called C3sh1B2herein), SEQ ID NO. 81/SEQ ID NO. 82 (called C3sh1B4 herein), SEQ ID NO.83/SEQ ID NO. 84 (called C3sh1B6 herein), SEQ ID NO. 85/SEQ ID NO. 86(called C3sh1B9 herein), SEQ ID NO. 87/SEQ ID NO. 88 (called C3sh1C1herein), SEQ ID NO. 89/SEQ ID NO. 90 (called C3sh1C2 herein), SEQ ID NO.91/SEQ ID NO. 92 (called C3sh1C7 herein), SEQ ID NO. 93/SEQ ID NO. 94(called C3sh1D1 herein), SEQ ID NO. 95/SEQ ID NO. 96 (called C3sh1D4herein), SEQ ID NO. 97/SEQ ID NO. 98 (called C3sh1D6 herein), SEQ ID NO.99/SEQ ID NO. 100 (called C3sh1E2 herein), SEQ ID NO. 101/SEQ ID NO. 102(called C3sh1E7 herein), SEQ ID NO. 103/SEQ ID NO. 104 (called C3sh1E9herein), SEQ ID NO. 105/SEQ ID NO. 106 (called C3sh1F1 herein), SEQ IDNO. 107/SEQ ID NO. 108 (called C3sh1F10 herein), SEQ ID NO. 109/SEQ IDNO. 110 (called C3sh1F12 herein), SEQ ID NO. 111/SEQ ID NO. 112 (calledC3sh1F2 herein), SEQ ID NO. 113/SEQ ID NO. 114 (called C3sh1G1 herein),SEQ ID NO. 115/SEQ ID NO. 116 (called C3sh1G11 herein), SEQ ID NO.117/SEQ ID NO. 118 (called C3sh1G2 herein), SEQ ID NO. 119/SEQ ID NO.120 (called C3sh1G3 herein), SEQ ID NO. 121/SEQ ID NO. 122 (calledC3sh1G5 herein), SEQ ID NO. 123/SEQ ID NO. 124 (called C3sh1G8 herein),SEQ ID NO. 125/SEQ ID NO. 126 (called C3sh1H10 herein), SEQ ID NO.127/SEQ ID NO. 128 (called C3sh1H4 herein), SEQ ID NO. 129/SEQ ID NO. 28(called MA8 herein), SEQ ID NO. 130/SEQ ID NO. 28 (called MB1 herein),SEQ ID NO. 131/SEQ ID NO. 28 (called MB3 herein), SEQ ID NO. 132/SEQ IDNO. 28 (called MB10 herein), SEQ ID NO. 133/SEQ ID NO. 28 (called MB12herein), SEQ ID NO. 134/SEQ ID NO. 28 (called MC8 herein), SEQ ID NO.135/SEQ ID NO. 28 (called MD1 herein), SEQ ID NO. 136/SEQ ID NO. 28(called MD4 herein), SEQ ID NO. 137/SEQ ID NO. 28 (called MSA11 herein),SEQ ID NO. 138/SEQ ID NO. 28 (called MSB7 herein), SEQ ID NO. 139/SEQ IDNO. 28 (called MSD2 herein), SEQ ID NO. 140/SEQ ID NO. 28 (called MSE3herein), SEQ ID NO. 141/SEQ ID NO. 28 (called MSE5 herein), SEQ ID NO.142/SEQ ID NO. 28 (called MSC8 herein), SEQ ID NO. 143/SEQ ID NO. 28(called MSH1 herein), and combinations thereof.

In one embodiment, the invention provides an anti-CD137 antibody, or anantigen-binding fragment thereof, comprising a heavy chain comprising aCDR3 domain as set forth in any one of SEQ ID Nos: 1, 3, 5, 7, 9, 11,13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47,49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83,85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115,117, 119, 121, 123, 125, 127, 129, 130, 131, 132, 133, 134, 135, 136,137, 139, 140, 141, 142 and 143 and comprising a variable domaincomprising an amino acid sequence that has at least 95%, at least 96%,at least 97%, at least 98%, or at least 99% identical to a sequence asset forth in any one of SEQ ID Nos: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19,21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55,57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91,93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121,123, 125, 127, 129, 130, 131, 132, 133, 134, 135, 136, 137, 139, 140,141, 142 and 143. In one embodiment, the invention provides ananti-CD137 antibody, or an antigen-binding fragment thereof, comprisinga light chain comprising a CDR3 domain as set forth in any one of SEQ IDNos: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36,38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72,74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106,108, 110, 112, 114, 116, 118, 120, 122, 124, 126 and 128 and having alight chain variable domain comprising an amino acid sequence that hasat least 95%, at least 96%, at least 97%, at least 98%, or at least 99%identical to a sequence as set forth in any one of SEQ ID Nos: 2, 4, 6,8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42,44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78,80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110,112, 114, 116, 118, 120, 122, 124, 126 and 128. Thus, in certainembodiments, the CDR3 domain is held constant, while variability may beintroduced into the remaining CDRs and/or framework regions of the heavyand/or light chains, while the antibody, or antigen binding fragmentthereof, retains the ability to bind to CD137 and retains the functionalcharacteristics, e.g., binding affinity, of the parent.

In one embodiment, the substitutions made within a heavy or light chainthat is at least 95% identical (or at least 96% identical, or at least97% identical, or at least 98% identical, or at least 99% identical) areconservative amino acid substitutions. A “conservative amino acidsubstitution” is one in which an amino acid residue is substituted byanother amino acid residue having a side chain (R group) with similarchemical properties (e.g., charge or hydrophobicity). In general, aconservative amino acid substitution will not substantially change thefunctional properties of a protein. In cases where two or more aminoacid sequences differ from each other by conservative substitutions, thepercent sequence identity or degree of similarity may be adjustedupwards to correct for the conservative nature of the substitution.Means for making this adjustment are well-known to those of skill in theart. See, e.g., Pearson (1994) Methods Mol. Biol. 24: 307-331, hereinincorporated by reference. Examples of groups of amino acids that haveside chains with similar chemical properties include (1) aliphatic sidechains: glycine, alanine, valine, leucine and isoleucine; (2)aliphatic-hydroxyl side chains: serine and threonine; (3)amide-containing side chains: asparagine and glutamine; (4) aromaticside chains: phenylalanine, tyrosine, and tryptophan; (5) basic sidechains: lysine, arginine, and histidine; (6) acidic side chains:aspartate and glutamate, and (7) sulfur-containing side chains arecysteine and methionine.

In one embodiment, the present invention is directed to an antibody, oran antigen binding fragment thereof, having the antigen binding regionsof any of the antibodies described in Table 5.

In one embodiment, the present invention is directed to an antibody, oran antigen binding fragment thereof, having antigen binding regions ofantibody D6. In one embodiment, the invention provides an antibody, orantigen-binding fragment thereof, comprising a heavy chain variabledomain sequence as set forth in SEQ ID NO: 27, and a light chainvariable domain sequence as set forth in SEQ ID NO: 28. In oneembodiment, the invention is directed to an antibody having a heavychain variable domain comprising the CDRs of SEQ ID NO: 27, and a lightchain variable domain comprising the CDRs of SEQ ID NO:28. In oneembodiment, the invention features an isolated human antibody, orantigen-binding fragment thereof, that comprises a heavy chain variableregion having an amino acid sequence that is at least 95% identical, atleast 96% identical, at least 97% identical, at least 98% identical, orat least 99% identical to the sequence set forth in SEQ ID NO: 27, andcomprises a light chain variable region having an amino acid sequencethat is at least 95% identical, at least 96% identical, at least 97%identical, at least 98% identical, or at least 99% identical to thesequence set forth in SEQ ID NO: 28. The antibody may further be an IgG1or an IgG4 isotype.

In one embodiment, the present invention is directed to an antibody, oran antigen binding fragment thereof, having antigen binding regions ofantibody MB3. In one embodiment, the invention provides an antibody, orantigen-binding fragment thereof, comprising a heavy chain variabledomain sequence as set forth in SEQ ID NO: 131, and a light chainvariable domain sequence as set forth in SEQ ID NO: 28. In oneembodiment, the invention is directed to an antibody having a heavychain variable domain comprising the CDRs of SEQ ID NO: 131, and a lightchain variable domain comprising the CDRs of SEQ ID NO:28. In oneembodiment, the invention features an isolated human antibody, orantigen-binding fragment thereof, that comprises a heavy chain variableregion having an amino acid sequence that is at least 95% identical, atleast 96% identical, at least 97% identical, at least 98% identical, orat least 99% identical to the sequence set forth in SEQ ID NO: 131 andcomprises a light chain variable region having an amino acid sequencethat is at least 95% identical, at least 96% identical, at least 97%identical, at least 98% identical, or at least 99% identical to thesequence set forth in SEQ ID NO:28. The antibody may further be an IgG1or an IgG4 isotype.

In one embodiment, the present invention is directed to an antibody, oran antigen binding fragment thereof, having antigen binding regions ofantibody MSH1. In one embodiment, the invention provides an antibody, orantigen-binding fragment thereof, comprising a heavy chain variabledomain sequence as set forth in SEQ ID NO: 143, and a light chainvariable domain sequence as set forth in SEQ ID NO: 28. In oneembodiment, the invention is directed to an antibody having a heavychain variable domain comprising the CDRs of SEQ ID NO: 143, and a lightchain variable domain comprising the CDRs of SEQ ID NO:28. In oneembodiment, the invention features an isolated human antibody, orantigen-binding fragment thereof, that comprises a heavy chain variableregion having an amino acid sequence that is at least 95% identical, atleast 96% identical, at least 97% identical, at least 98% identical, orat least 99% identical to the sequence set forth in SEQ ID NO: 143 andcomprises a light chain variable region having an amino acid sequencethat is at least 95% identical, at least 96% identical, at least 97%identical, at least 98% identical, or at least 99% identical to thesequence set forth in SEQ ID NO: 28. The antibody may further be an IgG1or an IgG4 isotype.

In one embodiment, the present invention is directed to an antibody, oran antigen binding fragment thereof, having antigen binding regions ofantibody MB12. In one embodiment, the invention provides an antibody, orantigen-binding fragment thereof, comprising a heavy chain variabledomain sequence as set forth in SEQ ID NO: 133, and a light chainvariable domain sequence as set forth in SEQ ID NO: 28. In oneembodiment, the invention is directed to an antibody having a heavychain variable domain comprising the CDRs of SEQ ID NO: 133, and a lightchain variable domain comprising the CDRs of SEQ ID NO:28. In oneembodiment, the invention features an isolated human antibody, orantigen-binding fragment thereof, that comprises a heavy chain variableregion having an amino acid sequence that is at least 95% identical, atleast 96% identical, at least 97% identical, at least 98% identical, orat least 99% identical to the sequence set forth in SEQ ID NO: 133 andcomprises a light chain variable region having an amino acid sequencethat is at least 95% identical, at least 96% identical, at least 97%identical, at least 98% identical, or at least 99% identical to thesequence set forth in SEQ ID NO: 28. The antibody may further be an IgG1or an IgG4 isotype.

In one embodiment, the present invention is directed to an antibody, oran antigen binding fragment thereof, having antigen binding regions ofantibody MB10. In one embodiment, the invention provides an antibody, orantigen-binding fragment thereof, comprising a heavy chain variabledomain sequence as set forth in SEQ ID NO: 132, and a light chainvariable domain sequence as set forth in SEQ ID NO: 28. In oneembodiment, the invention is directed to an antibody having a heavychain variable domain comprising the CDRs of SEQ ID NO: 132, and a lightchain variable domain comprising the CDRs of SEQ ID NO:28. In oneembodiment, the invention features an isolated human antibody, orantigen-binding fragment thereof, that comprises a heavy chain variableregion having an amino acid sequence that is at least 95% identical, atleast 96% identical, at least 97% identical, at least 98% identical, orat least 99% identical to the sequence set forth in SEQ ID NO: 132 andcomprises a light chain variable region having an amino acid sequencethat is at least 95% identical, at least 96% identical, at least 97%identical, at least 98% identical, or at least 99% identical to thesequence set forth in SEQ ID NO: 28. The antibody may further be an IgG1or an IgG4 isotype.

In one embodiment, the present invention is directed to an antibody, oran antigen binding fragment thereof, having antigen binding regions ofantibody MB1. In one embodiment, the invention provides an antibody, orantigen-binding fragment thereof, comprising a heavy chain variabledomain sequence as set forth in SEQ ID NO: 130, and a light chainvariable domain sequence as set forth in SEQ ID NO: 28. In oneembodiment, the invention is directed to an antibody having a heavychain variable domain comprising the CDRs of SEQ ID NO: 130, and a lightchain variable domain comprising the CDRs of SEQ ID NO:28. In oneembodiment, the invention features an isolated human antibody, orantigen-binding fragment thereof, that comprises a heavy chain variableregion having an amino acid sequence that is at least 95% identical, atleast 96% identical, at least 97% identical, at least 98% identical, orat least 99% identical to the sequence set forth in SEQ ID NO: 130 andcomprises a light chain variable region having an amino acid sequencethat is at least 95% identical, at least 96% identical, at least 97%identical, at least 98% identical, or at least 99% identical to thesequence set forth in SEQ ID NO: 28. The antibody may further be an IgG1or an IgG4 isotype.

In one embodiment, the present invention is directed to an antibody, oran antigen binding fragment thereof, having antigen binding regions ofantibody MSC8. In one embodiment, the invention provides an antibody, orantigen-binding fragment thereof, comprising a heavy chain variabledomain sequence as set forth in SEQ ID NO: 142, and a light chainvariable domain sequence as set forth in SEQ ID NO: 28. In oneembodiment, the invention is directed to an antibody having a heavychain variable domain comprising the CDRs of SEQ ID NO: 142, and a lightchain variable domain comprising the CDRs of SEQ ID NO:28. In oneembodiment, the invention features an isolated human antibody, orantigen-binding fragment thereof, that comprises a heavy chain variableregion having an amino acid sequence that is at least 95% identical, atleast 96% identical, at least 97% identical, at least 98% identical, orat least 99% identical to the sequence set forth in SEQ ID NO: 142 andcomprises a light chain variable region having an amino acid sequencethat is at least 95% identical, at least 96% identical, at least 97%identical, at least 98% identical, or at least 99% identical to thesequence set forth in SEQ ID NO: 28. The antibody may further be an IgG1or an IgG4 isotype.

In one embodiment, the present invention is directed to an antibody, oran antigen binding fragment thereof, having antigen binding regions ofantibody MSB7. In one embodiment, the invention provides an antibody, orantigen-binding fragment thereof, comprising a heavy chain variabledomain sequence as set forth in SEQ ID NO: 138, and a light chainvariable domain sequence as set forth in SEQ ID NO: 28. In oneembodiment, the invention is directed to an antibody having a heavychain variable domain comprising the CDRs of SEQ ID NO: 138, and a lightchain variable domain comprising the CDRs of SEQ ID NO:28. The antibodymay further be an IgG1 or an IgG4 isotype.

As described in Table 5, a number of heavy chain variable domain aminoacid sequences are at least 95% identical to SEQ ID NO: 27. A number ofheavy chain variable domains have amino acid sequences that are at least95% identical to SEQ ID NO:27, including SEQ ID NO: 143 (as describedfor antibody MSH1), SEQ ID NO: 142 (as described for antibody MSC8), SEQID NO: 130 (as described for antibody MB1), SEQ ID NO: 131 (as describedfor antibody MB3), SEQ ID NO: 132 (as described for antibody MB10), SEQID NO: 129 (as described for antibody MA8), SEQ ID NO: 133 (as describedfor antibody MB12) and SEQ ID NO: 135 (as described for antibody MD1).

A number of heavy chain variable domains have amino acid sequences thatare at least 95% identical to SEQ ID NO:141, including SEQ ID NO: 139(as described for antibody MSD2), SEQ ID NO: 143 (as described forantibody MSH1), SEQ ID NO: 142 (as described for antibody MSC8) and SEQID NO: 129 (as described for antibody MA8).

A number of heavy chain variable domains have amino acid sequences thatare at least 95% identical to SEQ ID NO:139, including SEQ ID NO: 143(as described for antibody MSH1) and SEQ ID NO: 142 (as described forantibody MSC8).).

A number of heavy chain variable domains have amino acid sequences thatare at least 95% identical to SEQ ID NO:143, including SEQ ID NO: 139(as described for antibody MSD2), SEQ ID NO: 142 (as described forantibody MSC8), SEQ ID NO: 130 (as described for antibody MB1), SEQ IDNO: 27 (as described for antibody D6) and SEQ ID NO: 129 (as describedfor antibody MA8).

A number of heavy chain variable domains have amino acid sequences thatare at least 95% identical to SEQ ID NO:142, including SEQ ID NO: 141(as described for antibody MSE5), SEQ ID NO: 139 (as described forantibody MSD2), SEQ ID NO: 142 (as described for antibody MSC8), SEQ IDNO: 143 (as described for antibody MSH1), SEQ ID NO: 130 (as describedfor antibody MB1), SEQ ID NO: 131 (as described for antibody MB3), SEQID NO: 132 (as described for antibody MB10), SEQ ID NO: 27 (as describedfor antibody D6), SEQ ID NO: 133 (as described for antibody MB12) andSEQ ID NO: 135 (as described for antibody MD1).

A number of heavy chain variable domains have amino acid sequences thatare at least 95% identical to SEQ ID NO:130, including SEQ ID NO: 143(as described for antibody MSH1), SEQ ID NO: 142 (as described forantibody MSC8), SEQ ID NO: 131 (as described for antibody MB3), SEQ IDNO: 132 (as described for antibody MB10), SEQ ID NO: 27 (as describedfor antibody D6), SEQ ID NO: 133 (as described for antibody MB12) andSEQ ID NO: 135 (as described for antibody MD1).

A number of heavy chain variable domains have amino acid sequences thatare at least 95% identical to SEQ ID NO:131, including SEQ ID NO: 142(as described for antibody MSC8), SEQ ID NO: 130 (as described forantibody MB1), SEQ ID NO: 132 (as described for antibody MB10), SEQ IDNO: 27 (as described for antibody D6), SEQ ID NO: 133 (as described forantibody MB12) and SEQ ID NO: 135 (as described for antibody MD1).

A number of heavy chain variable domains have amino acid sequences thatare at least 95% identical to SEQ ID NO:132, including SEQ ID NO: 142(as described for antibody MSC8), SEQ ID NO: 130 (as described forantibody MB1), SEQ ID NO: 131 (as described for antibody MB3), SEQ IDNO: 132 (as described for antibody MB10), SEQ ID NO: 27 (as describedfor antibody D6), SEQ ID NO: 133 (as described for antibody MB12) andSEQ ID NO: 135 (as described for antibody MD1).

A number of heavy chain variable domains have amino acid sequences thatare at least 95% identical to SEQ ID NO:129, including SEQ ID NO: 143(as described for antibody MSH1), SEQ ID NO: 130 (as described forantibody MB1), SEQ ID NO: 131 (as described for antibody MB3), SEQ IDNO: 132 (as described for antibody MB10), SEQ ID NO: 27 (as describedfor antibody D6), SEQ ID NO: 133 (as described for antibody MB12) andSEQ ID NO: 135 (as described for antibody MD1).

A number of heavy chain variable domains have amino acid sequences thatare at least 95% identical to SEQ ID NO:133, including SEQ ID NO: 142(as described for antibody MSC8), SEQ ID NO: 130 (as described forantibody MB1), SEQ ID NO: 131 (as described for antibody MB3), SEQ IDNO: 132 (as described for antibody MB10), SEQ ID NO: 27 (as describedfor antibody D6), SEQ ID NO: 129 (as described for antibody MA8) and SEQID NO: 135 (as described for antibody MD1).

A number of heavy chain variable domains have amino acid sequences thatare at least 95% identical to SEQ ID NO:135, including SEQ ID NO: 142(as described for antibody MSC8), SEQ ID NO: 130 (as described forantibody MB1), SEQ ID NO: 131 (as described for antibody MB3), SEQ IDNO: 132 (as described for antibody MB10), SEQ ID NO: 27 (as describedfor antibody D6) and SEQ ID NO: 133 (as described for antibody MB12).

SEQ ID NO 28 is included as the light chain variable domain in a numberof antibodies, including D6, MA8, MB1, MB3, MB10, MB12, MC8, MD1, MD4,MSA11, MSB7, MSD2, MSE3, MSE5, MSC8 and MSH1, as described in Table 5. Anumber of light chain variable domains have amino acid sequences thatare at least 95% identical to SEQ ID NO:28, including SEQ ID NO: 62 (asdescribed for antibody H4), SEQ ID NO: 90 (as described for antibodyC3sh1C2) and SEQ ID NO: 84 (as described for antibody C3sh1B6).

A number of light chain variable domains have amino acid sequences thatare at least 95% identical to SEQ ID NO:22, including SEQ ID NO: 118 (asdescribed for antibody C3sh1G2) and SEQ ID NO: 14 (as described forantibody C2).

A number of light chain variable domains have amino acid sequences thatare at least 95% identical to SEQ ID NO:128, including SEQ ID NO: 50 (asdescribed for antibody G2) and SEQ ID NO: 126 (as described for antibodyC3sh1H10).

A number of light chain variable domains have amino acid sequences thatare at least 95% identical to SEQ ID NO:126, including SEQ ID NO: 50 (asdescribed for antibody G2), SEQ ID NO: 128 (as described for antibodyC3sh1H4), SEQ ID NO:40 (as described for antibody E7), and SEQ ID NO: 98(as described for antibody C3sh1D6).

Antigen binding proteins (e.g., antibodies, antibody fragments, antibodyderivatives, antibody muteins, and antibody variants) are polypeptidesthat bind to CD137.

Antigen-binding fragments of antigen binding proteins of the inventionmay be produced by conventional techniques. Examples of such fragmentsinclude, but are not limited to, Fab and F(ab′)2 fragments.

Single chain antibodies may be formed by linking heavy and light chainvariable domain (Fv region) fragments via an amino acid bridge (shortpeptide linker), resulting in a single polypeptide chain. Suchsingle-chain Fvs (scFvs) have been prepared by fusing DNA encoding apeptide linker between DNAs encoding the two variable domainpolypeptides (VL and VH). The resulting polypeptides can fold back onthemselves to form antigen-binding monomers, or they can form multimers(e.g., dimers, trimers, or tetramers), depending on the length of aflexible linker between the two variable domains (Kortt et al., 1997,Prot. Eng. 10:423; Kortt et al., 2001, Biomol. Eng. 18:95-108). Bycombining different VL and VH-comprising polypeptides, one can formmultimeric scFvs that bind to different epitopes (Kriangkum et al.,2001, Biomol. Eng. 18:31-40). Techniques developed for the production ofsingle chain antibodies include those described in U.S. Pat. No.4,946,778; Bird, 1988, Science 242:423; Huston et al., 1988, Proc. Natl.Acad. Sci. USA 85:5879; Ward et al., 1989, Nature 334:544, de Graaf etal., 2002, Methods Mol. Biol. 178:379-87.

In certain embodiments, the present disclosure provides a Fab fullyhuman antibody fragment, having a variable domain region from a heavychain and a variable domain region from a light chain, wherein the heavychain variable domain sequence that is at least 95% identical, at least96% identical, at least 97% identical, at least 98% identical, or atleast 99% identical, to the amino acid sequences selected from the groupconsisting of SEQ ID NO. 1, SEQ ID NO. 3, SEQ ID NO. 5, SEQ ID NO. 6,SEQ ID NO. 7, SEQ ID NO. 9, SEQ ID NO. 11, SEQ ID NO. 13, SEQ ID NO. 15,SEQ ID NO. 17, SEQ ID NO. 19, SEQ ID NO. 21, SEQ ID NO. 23, SEQ ID NO.25, SEQ ID NO. 27, SEQ ID NO. 29, SEQ ID NO. 31, SEQ ID NO. 33, SEQ IDNO. 35, SEQ ID NO. 37, SEQ ID NO. 39, SEQ ID NO. 41, SEQ ID NO. 43, SEQID NO. 45, SEQ ID NO. 47, SEQ ID NO. 49, SEQ ID NO. 51, SEQ ID NO. 53,SEQ ID NO. 55, SEQ ID NO. 57, SEQ ID NO. 59, SEQ ID NO. 61, SEQ ID NO.63, SEQ ID NO. 65, SEQ ID NO. 67, SEQ ID NO. 69, SEQ ID NO. 71, SEQ IDNO. 73, SEQ ID NO. 75, SEQ ID NO. 77, SEQ ID NO. 79, SEQ ID NO. 81, SEQID NO. 83, SEQ ID NO. 85, SEQ ID NO. 87, SEQ ID NO. 89, SEQ ID NO. 91,SEQ ID NO. 93, SEQ ID NO. 95, SEQ ID NO. 97, SEQ ID NO. 99, SEQ ID NO.101, SEQ ID NO. 103, SEQ ID NO. 105, SEQ ID NO. 107, SEQ ID NO. 109, SEQID NO. 111, SEQ ID NO. 113, SEQ ID NO. 115, SEQ ID NO. 117, SEQ ID NO.119, SEQ ID NO. 121, SEQ ID NO. 123, SEQ ID NO. 125, SEQ ID NO. 127, SEQID NO. 129, SEQ ID NO. 130, SEQ ID NO. 131, SEQ ID NO. 132, SEQ ID NO.133, SEQ ID NO. 134, SEQ ID NO. 135, SEQ ID NO. 136, SEQ ID NO. 137, SEQID NO. 138, SEQ ID NO. 139, SEQ ID NO. 140, SEQ ID NO. 141, SEQ ID NO.142, SEQ ID NO. 143, and combinations thereof, and that has a lightchain variable domain sequence that is at least 95% identical, at least96% identical, at least 97% identical, at least 98% identical, or atleast 99% identical, to the amino acid sequence consisting of SEQ ID NO.2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO.12, SEQ ID NO. 14, SEQ ID NO. 16, SEQ ID NO. 18, SEQ ID NO. 20, SEQ IDNO. 22, SEQ ID NO. 24, SEQ ID NO. 26, SEQ ID NO. 28, SEQ ID NO. 30, SEQID NO. 32, SEQ ID NO. 34, SEQ ID NO. 36, SEQ ID NO. 38, SEQ ID NO. 40,SEQ ID NO. 42, SEQ ID NO. 44, SEQ ID NO. 46, SEQ ID NO. 48, SEQ ID NO.50, SEQ ID NO. 52, SEQ ID NO. 54, SEQ ID NO. 56, SEQ ID NO. 58, SEQ IDNO. 60, SEQ ID NO. 62, SEQ ID NO. 64, SEQ ID NO. 66, SEQ ID NO. 68, SEQID NO. 70, SEQ ID NO. 72, SEQ ID NO. 74, SEQ ID NO. 76, SEQ ID NO. 78,SEQ ID NO. 80, SEQ ID NO. 82, SEQ ID NO. 84, SEQ ID NO. 86, SEQ ID NO.88, SEQ ID NO. 90, SEQ ID NO. 92, SEQ ID NO. 94, SEQ ID NO. 96, SEQ IDNO. 98, SEQ ID NO. 100, SEQ ID NO. 102, SEQ ID NO. 104, SEQ ID NO. 106,SEQ ID NO. 108, SEQ ID NO. 110, SEQ ID NO. 112, SEQ ID NO. 114, SEQ IDNO. 116, SEQ ID NO. 118, SEQ ID NO. 120, SEQ ID NO. 122, SEQ ID NO. 124,SEQ ID NO. 126, SEQ ID NO. 128, and combinations thereof. Preferably,the fully human antibody Fab fragment has both a heavy chain variabledomain region and a light chain variable domain region wherein theantibody has a heavy chain/light chain variable domain sequence selectedfrom the group consisting of SEQ ID NO. 1/SEQ ID NO. 2, SEQ ID NO. 3/SEQID NO. 4, SEQ ID NO. 5/SEQ ID NO. 6, SEQ ID NO. 7/SEQ ID NO. 8, SEQ IDNO. 9/SEQ ID NO. 10, SEQ ID NO. 11/SEQ ID NO. 12, SEQ ID NO. 13/SEQ IDNO. 14, SEQ ID NO. 15/SEQ ID NO. 16, SEQ ID NO. 17/SEQ ID NO. 18, SEQ IDNO. 19/SEQ ID NO. 20, SEQ ID NO. 21/SEQ ID NO. 22, SEQ ID NO. 23/SEQ IDNO. 24, SEQ ID NO. 25/SEQ ID NO. 26, SEQ ID NO. 27/SEQ ID NO. 28, SEQ IDNO. 29/SEQ ID NO. 30, SEQ ID NO. 31/SEQ ID NO. 32, SEQ ID NO. 33/SEQ IDNO. 34, SEQ ID NO. 35/SEQ ID NO. 36, SEQ ID NO. 37/SEQ ID NO. 38, SEQ IDNO. 39/SEQ ID NO. 40, SEQ ID NO. 41/SEQ ID NO. 42, SEQ ID NO. 43/SEQ IDNO. 44, SEQ ID NO. 45/SEQ ID NO. 46, SEQ ID NO. 47/SEQ ID NO. 48, SEQ IDNO. 49/SEQ ID NO. 50, SEQ ID NO. 51/SEQ ID NO. 52, SEQ ID NO. 53/SEQ IDNO. 54, SEQ ID NO. 55/SEQ ID NO. 56, SEQ ID NO. 57/SEQ ID NO. 58, SEQ IDNO. 59/SEQ ID NO. 60, SEQ ID NO. 61/SEQ ID NO. 62, SEQ ID NO. 63/SEQ IDNO. 64, SEQ ID NO. 65/SEQ ID NO. 66, SEQ ID NO. 67/SEQ ID NO. 68, SEQ IDNO. 69/SEQ ID NO. 70, SEQ ID NO. 71/SEQ ID NO. 72, SEQ ID NO. 73/SEQ IDNO. 74, SEQ ID NO. 75/SEQ ID NO. 76, SEQ ID NO. 77/SEQ ID NO. 78, SEQ IDNO. 79/SEQ ID NO. 80, SEQ ID NO. 81/SEQ ID NO. 82, SEQ ID NO. 83/SEQ IDNO. 84, SEQ ID NO. 85/SEQ ID NO. 86, SEQ ID NO. 87/SEQ ID NO. 88, SEQ IDNO. 89/SEQ ID NO. 90, SEQ ID NO. 91/SEQ ID NO. 92, SEQ ID NO. 93/SEQ IDNO. 94, SEQ ID NO. 95/SEQ ID NO. 96, SEQ ID NO. 97/SEQ ID NO. 98, SEQ IDNO. 99/SEQ ID NO. 100, SEQ ID NO. 101/SEQ ID NO. 102, SEQ ID NO. 103/SEQID NO. 104, SEQ ID NO. 105/SEQ ID NO. 106, SEQ ID NO. 107/SEQ ID NO.108, SEQ ID NO. 109/SEQ ID NO. 110, SEQ ID NO. 111/SEQ ID NO. 112, SEQID NO. 113/SEQ ID NO. 114, SEQ ID NO. 115/SEQ ID NO. 116, SEQ ID NO.117/SEQ ID NO. 118, SEQ ID NO. 119/SEQ ID NO. 120, SEQ ID NO. 121/SEQ IDNO. 122, SEQ ID NO. 123/SEQ ID NO. 124, SEQ ID NO. 125/SEQ ID NO. 126,SEQ ID NO. 127/SEQ ID NO. 128, SEQ ID NO. 129/SEQ ID NO. 28, SEQ ID NO.130/SEQ ID NO. 28, SEQ ID NO. 131/SEQ ID NO. 28, SEQ ID NO. 132/SEQ IDNO. 28, SEQ ID NO. 133/SEQ ID NO. 28, SEQ ID NO. 134/SEQ ID NO. 28, SEQID NO. 135/SEQ ID NO. 28, SEQ ID NO. 136/SEQ ID NO. 28, SEQ ID NO.137/SEQ ID NO. 28, SEQ ID NO. 138/SEQ ID NO. 28, SEQ ID NO. 139/SEQ IDNO. 28, SEQ ID NO. 140/SEQ ID NO. 28, SEQ ID NO. 141/SEQ ID NO. 28, SEQID NO. 142/SEQ ID NO. 28, SEQ ID NO. 143/SEQ ID NO. 28, and combinationsthereof.

In one embodiment, the present disclosure provides a single chain humanantibody, having a variable domain region from a heavy chain and avariable domain region from a light chain and a peptide linkerconnection the heavy chain and light chain variable domain regions,wherein the heavy chain variable domain sequence that is at least 95%identical, at least 96% identical, at least 97% identical, at least 98%identical, or at least 99% identical, to the amino acid sequencesselected from the group consisting of SEQ ID NO. 1, SEQ ID NO. 3, SEQ IDNO. 5, SEQ ID NO. 6, SEQ ID NO. 7, SEQ ID NO. 9, SEQ ID NO. 11, SEQ IDNO. 13, SEQ ID NO. 15, SEQ ID NO. 17, SEQ ID NO. 19, SEQ ID NO. 21, SEQID NO. 23, SEQ ID NO. 25, SEQ ID NO. 27, SEQ ID NO. 29, SEQ ID NO. 31,SEQ ID NO. 33, SEQ ID NO. 35, SEQ ID NO. 37, SEQ ID NO. 39, SEQ ID NO.41, SEQ ID NO. 43, SEQ ID NO. 45, SEQ ID NO. 47, SEQ ID NO. 49, SEQ IDNO. 51, SEQ ID NO. 53, SEQ ID NO. 55, SEQ ID NO. 57, SEQ ID NO. 59, SEQID NO. 61, SEQ ID NO. 63, SEQ ID NO. 65, SEQ ID NO. 67, SEQ ID NO. 69,SEQ ID NO. 71, SEQ ID NO. 73, SEQ ID NO. 75, SEQ ID NO. 77, SEQ ID NO.79, SEQ ID NO. 81, SEQ ID NO. 83, SEQ ID NO. 85, SEQ ID NO. 87, SEQ IDNO. 89, SEQ ID NO. 91, SEQ ID NO. 93, SEQ ID NO. 95, SEQ ID NO. 97, SEQID NO. 99, SEQ ID NO. 101, SEQ ID NO. 103, SEQ ID NO. 105, SEQ ID NO.107, SEQ ID NO. 109, SEQ ID NO. 111, SEQ ID NO. 113, SEQ ID NO. 115, SEQID NO. 117, SEQ ID NO. 119, SEQ ID NO. 121, SEQ ID NO. 123, SEQ ID NO.125, SEQ ID NO. 127, SEQ ID NO. 129, SEQ ID NO. 130, SEQ ID NO. 131, SEQID NO. 132, SEQ ID NO. 133, SEQ ID NO. 134, SEQ ID NO. 135, SEQ ID NO.136, SEQ ID NO. 137, SEQ ID NO. 138, SEQ ID NO. 139, SEQ ID NO. 140, SEQID NO. 141, SEQ ID NO. 142, SEQ ID NO. 143, and combinations thereof,and that has a light chain variable domain sequence that is at least 95%identical, at least 96% identical, at least 97% identical, at least 98%identical, or at least 99% identical, to the amino acid sequenceconsisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8,SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, SEQ ID NO. 16, SEQ ID NO.18, SEQ ID NO. 20, SEQ ID NO. 22, SEQ ID NO. 24, SEQ ID NO. 26, SEQ IDNO. 28, SEQ ID NO. 30, SEQ ID NO. 32, SEQ ID NO. 34, SEQ ID NO. 36, SEQID NO. 38, SEQ ID NO. 40, SEQ ID NO. 42, SEQ ID NO. 44, SEQ ID NO. 46,SEQ ID NO. 48, SEQ ID NO. 50, SEQ ID NO. 52, SEQ ID NO. 54, SEQ ID NO.56, SEQ ID NO. 58, SEQ ID NO. 60, SEQ ID NO. 62, SEQ ID NO. 64, SEQ IDNO. 66, SEQ ID NO. 68, SEQ ID NO. 70, SEQ ID NO. 72, SEQ ID NO. 74, SEQID NO. 76, SEQ ID NO. 78, SEQ ID NO. 80, SEQ ID NO. 82, SEQ ID NO. 84,SEQ ID NO. 86, SEQ ID NO. 88, SEQ ID NO. 90, SEQ ID NO. 92, SEQ ID NO.94, SEQ ID NO. 96, SEQ ID NO. 98, SEQ ID NO. 100, SEQ ID NO. 102, SEQ IDNO. 104, SEQ ID NO. 106, SEQ ID NO. 108, SEQ ID NO. 110, SEQ ID NO. 112,SEQ ID NO. 114, SEQ ID NO. 116, SEQ ID NO. 118, SEQ ID NO. 120, SEQ IDNO. 122, SEQ ID NO. 124, SEQ ID NO. 126, SEQ ID NO. 128, andcombinations thereof. Preferably, the fully human single chain antibodyhas both a heavy chain variable domain region and a light chain variabledomain region, wherein the single chain fully human antibody has a heavychain/light chain variable domain sequence selected from the groupconsisting of SEQ ID NO. 1/SEQ ID NO. 2, SEQ ID NO. 3/SEQ ID NO. 4, SEQID NO. 5/SEQ ID NO. 6, SEQ ID NO. 7/SEQ ID NO. 8, SEQ ID NO. 9/SEQ IDNO. 10, SEQ ID NO. 11/SEQ ID NO. 12, SEQ ID NO. 13/SEQ ID NO. 14, SEQ IDNO. 15/SEQ ID NO. 16, SEQ ID NO. 17/SEQ ID NO. 18, SEQ ID NO. 19/SEQ IDNO. 20, SEQ ID NO. 21/SEQ ID NO. 22, SEQ ID NO. 23/SEQ ID NO. 24, SEQ IDNO. 25/SEQ ID NO. 26, SEQ ID NO. 27/SEQ ID NO. 28, SEQ ID NO. 29/SEQ IDNO. 30, SEQ ID NO. 31/SEQ ID NO. 32, SEQ ID NO. 33/SEQ ID NO. 34, SEQ IDNO. 35/SEQ ID NO. 36, SEQ ID NO. 37/SEQ ID NO. 38, SEQ ID NO. 39/SEQ IDNO. 40, SEQ ID NO. 41/SEQ ID NO. 42, SEQ ID NO. 43/SEQ ID NO. 44, SEQ IDNO. 45/SEQ ID NO. 46, SEQ ID NO. 47/SEQ ID NO. 48, SEQ ID NO. 49/SEQ IDNO. 50, SEQ ID NO. 51/SEQ ID NO. 52, SEQ ID NO. 53/SEQ ID NO. 54, SEQ IDNO. 55/SEQ ID NO. 56, SEQ ID NO. 57/SEQ ID NO. 58, SEQ ID NO. 59/SEQ IDNO. 60, SEQ ID NO. 61/SEQ ID NO. 62, SEQ ID NO. 63/SEQ ID NO. 64, SEQ IDNO. 65/SEQ ID NO. 66, SEQ ID NO. 67/SEQ ID NO. 68, SEQ ID NO. 69/SEQ IDNO. 70, SEQ ID NO. 71/SEQ ID NO. 72, SEQ ID NO. 73/SEQ ID NO. 74, SEQ IDNO. 75/SEQ ID NO. 76, SEQ ID NO. 77/SEQ ID NO. 78, SEQ ID NO. 79/SEQ IDNO. 80, SEQ ID NO. 81/SEQ ID NO. 82, SEQ ID NO. 83/SEQ ID NO. 84, SEQ IDNO. 85/SEQ ID NO. 86, SEQ ID NO. 87/SEQ ID NO. 88, SEQ ID NO. 89/SEQ IDNO. 90, SEQ ID NO. 91/SEQ ID NO. 92, SEQ ID NO. 93/SEQ ID NO. 94, SEQ IDNO. 95/SEQ ID NO. 96, SEQ ID NO. 97/SEQ ID NO. 98, SEQ ID NO. 99/SEQ IDNO. 100, SEQ ID NO. 101/SEQ ID NO. 102, SEQ ID NO. 103/SEQ ID NO. 104,SEQ ID NO. 105/SEQ ID NO. 106, SEQ ID NO. 107/SEQ ID NO. 108, SEQ ID NO.109/SEQ ID NO. 110, SEQ ID NO. 111/SEQ ID NO. 112, SEQ ID NO. 113/SEQ IDNO. 114, SEQ ID NO. 115/SEQ ID NO. 116, SEQ ID NO. 117/SEQ ID NO. 118,SEQ ID NO. 119/SEQ ID NO. 120, SEQ ID NO. 121/SEQ ID NO. 122, SEQ ID NO.123/SEQ ID NO. 124, SEQ ID NO. 125/SEQ ID NO. 126, SEQ ID NO. 127/SEQ IDNO. 128, SEQ ID NO. 129/SEQ ID NO. 28, SEQ ID NO. 130/SEQ ID NO. 28, SEQID NO. 131/SEQ ID NO. 28, SEQ ID NO. 132/SEQ ID NO. 28, SEQ ID NO.133/SEQ ID NO. 28, SEQ ID NO. 134/SEQ ID NO. 28, SEQ ID NO. 135/SEQ IDNO. 28, SEQ ID NO. 136/SEQ ID NO. 28, SEQ ID NO. 137/SEQ ID NO. 28, SEQID NO. 138/SEQ ID NO. 28, SEQ ID NO. 139/SEQ ID NO. 28, SEQ ID NO.140/SEQ ID NO. 28, SEQ ID NO. 141/SEQ ID NO. 28, SEQ ID NO. 142/SEQ IDNO. 28, SEQ ID NO. 143/SEQ ID NO. 28, and combinations thereof.

Techniques are known for deriving an antibody of a different subclass orisotype from an antibody of interest, i.e., subclass switching. Thus,IgG antibodies may be derived from an IgM antibody, for example, andvice versa. Such techniques allow the preparation of new antibodies thatpossess the antigen-binding properties of a given antibody (the parentantibody), but also exhibit biological properties associated with anantibody isotype or subclass different from that of the parent antibody.Recombinant DNA techniques may be employed. Cloned DNA encodingparticular antibody polypeptides may be employed in such procedures,e.g., DNA encoding the constant domain of an antibody of the desiredisotype (Lantto et al., 2002, Methods Mol. Biol. 178:303-16). Moreover,if an IgG4 is desired, it may also be desired to introduce a pointmutation (CPSC->CPPC) in the hinge region (Bloom et al., 1997, ProteinScience 6:407) to alleviate a tendency to form intra-H chain disulfidebonds that can lead to heterogeneity in the IgG4 antibodies. Thus, inone embodiment, the antibody of the invention is a human IgG1 antibody.Thus, in one embodiment, the antibody of the invention is a human IgG4antibody.

The present disclosure provides a number of antibodies structurallycharacterized by the amino acid sequences of their variable domainregions. However, the amino acid sequences can undergo some changeswhile retaining their high degree of binding to their specific targets.More specifically, many amino acids in the variable domain region can bechanged with conservative substitutions and it is predictable that thebinding characteristics of the resulting antibody will not differ fromthe binding characteristics of the wild type antibody sequence. Thereare many amino acids in an antibody variable domain that do not directlyinteract with the antigen or impact antigen binding and are not criticalfor determining antibody structure. For example, a predictednonessential amino acid residue in any of the disclosed antibodies ispreferably replaced with another amino acid residue from the same class.Methods of identifying amino acid conservative substitutions which donot eliminate antigen binding are well-known in the art (see, e.g.,Brummell et al., Biochem. 32: 1180-1187 (1993); Kobayashi et al. ProteinEng. 12(10):879-884 (1999); and Burks et al. Proc. Natl. Acad. Sci. USA94:412-417 (1997)). Near et al. Mol. Immunol. 30:369-377, 1993 explainshow to impact or not impact binding through site-directed mutagenesis.Near et al. only mutated residues that they thought had a highprobability of changing antigen binding. Most had a modest or negativeeffect on binding affinity (Near et al. Table 3) and binding todifferent forms of digoxin (Near et al. Table 2). Thus, the inventionalso includes, in certain embodiments, variable sequences having atleast 95% identity, at least 96% identity, at least 97% identity, atleast 98% identity, or at least 99% identity to those sequencesdisclosed herein.

In certain embodiments, an antibody, or antigen-binding fragmentthereof, provided herein has a dissociation constant (K_(d)) of 1×10⁻⁶ Mor less; 5×10⁻⁷ M or less 1×10⁻⁷ M or less; 5'10⁻⁸ M or less; 1×10⁻⁸ Mor less; 5×10⁻⁹ M or less; or 1×10⁻⁹ M or less. In one embodiment, theantibody, or antigen-binding fragment thereof, of the invention as a Kdfrom 1×10⁻⁷ M to 1×10⁻¹⁰ M. In one embodiment, the antibody, orantigen-binding fragment thereof, of the invention as a K_(d) from1×10⁻⁸ M to 1×10⁻¹⁰ M.

Those of ordinary skill in the art will appreciate standard methodsknown for determining the Kd of an antibody, or fragment thereof. Forexample, in one embodiment, Kd is measured by a radiolabeled antigenbinding assay (RIA). In one embodiment, an RIA is performed with the Fabversion of an antibody of interest and its antigen. For example,solution binding affinity of Fabs for antigen is measured byequilibrating Fab with a minimal concentration of (¹²⁵I)-labeled antigenin the presence of a titration series of unlabeled antigen, thencapturing bound antigen with an anti-Fab antibody-coated plate (see,e.g., Chen et al., J. Mol. Biol. 293:865-881(1999)).

According to another embodiment, Kd is measured using a BIACORE surfaceplasmon resonance assay. The term “surface plasmon resonance”, as usedherein, refers to an optical phenomenon that allows for the analysis ofreal-time interactions by detection of alterations in proteinconcentrations within a biosensor matrix, for example using the BIAcore™system (Biacore Life Sciences division of GE Healthcare, Piscataway,N.J.).

In particular embodiments, antigen binding proteins of the presentinvention have a binding affinity (K_(a)) for CD137 of at least 10⁶. Inother embodiments, the antigen binding proteins exhibit a K_(a) of atleast 10⁷, at least 10⁸, at least 10⁹, or at least 10¹⁰. In anotherembodiment, the antigen binding protein exhibits a K_(a) substantiallythe same as that of an antibody described herein in the Examples.

In another embodiment, the present disclosure provides an antigenbinding protein that has a low dissociation rate from CD137. In oneembodiment, the antigen binding protein has a K_(off) of 1×10⁻⁴ to ⁻¹ orlower. In another embodiment, the K_(off) is 5×10⁻⁵ to ⁻¹ or lower. Inanother embodiment, the K_(off) is substantially the same as an antibodydescribed herein. In another embodiment, the antigen binding proteinbinds to CD137 with substantially the same K_(off) as an antibodydescribed herein.

In another aspect, the present disclosure provides an antigen bindingprotein that binds to CD137 expressed on the surface of a cell and, whenso bound, inhibits CD137 signaling activity in the cell without causinga significant reduction in the amount of CD137 on the surface of thecell. Any method for determining or estimating the amount of CD137 onthe surface and/or in the interior of the cell can be used. In otherembodiments, binding of the antigen binding protein to theCD137-expressing cell causes less than about 75%, 50%, 40%, 30%, 20%,15%, 10%, 5%, 1%, or 0.1% of the cell-surface CD137 to be internalized.

In another aspect, the present disclosure provides an antigen bindingprotein having a half-life of at least one day in vitro or in vivo(e.g., when administered to a human subject). In one embodiment, theantigen binding protein has a half-life of at least three days. Inanother embodiment, the antigen binding protein has a half-life of fourdays or longer. In another embodiment, the antigen binding protein has ahalf-life of eight days or longer. In another embodiment, the antigenbinding protein is derivatized or modified such that it has a longerhalf-life as compared to the underivatized or unmodified antigen bindingprotein. In another embodiment, the antigen binding protein contains oneor more point mutations to increase serum half-life, such as describedin WO00/09560, incorporated by reference herein.

The present disclosure further provides multi-specific antigen bindingproteins, for example, bispecific antigen binding protein, e.g., antigenbinding protein that bind to two different epitopes of CD137, or to anepitope of CD137 and an epitope of another molecule, via two differentantigen binding sites or regions. Moreover, bispecific antigen bindingprotein as disclosed herein can comprise a CD137 binding site from oneof the herein-described antibodies and a second CD137 binding regionfrom another of the herein-described antibodies, including thosedescribed herein by reference to other publications. Alternatively, abispecific antigen binding protein may comprise an antigen binding sitefrom one of the herein described antibodies and a second antigen bindingsite from another CD137 antibody that is known in the art, or from anantibody that is prepared by known methods or the methods describedherein.

Numerous methods of preparing bispecific antibodies are known in theart. Such methods include the use of hybrid-hybridomas as described byMilstein et al., 1983, Nature 305:537, and chemical coupling of antibodyfragments (Brennan et al., 1985, Science 229:81; Glennie et al., 1987,J. Immunol. 139:2367; U.S. Pat. No. 6,010,902). Moreover, bispecificantibodies can be produced via recombinant means, for example by usingleucine zipper moieties (i.e., from the Fos and Jun proteins, whichpreferentially form heterodimers; Kostelny et al., 1992, J. Immunol.148:1547) or other lock and key interactive domain structures asdescribed in U.S. Pat. No. 5,582,996. Additional useful techniquesinclude those described in U.S. Pat. Nos. 5,959,083; and 5,807,706.

In another aspect, the antigen binding protein comprises a derivative ofan antibody. The derivatized antibody can comprise any molecule orsubstance that imparts a desired property to the antibody, such asincreased half-life in a particular use. The derivatized antibody cancomprise, for example, a detectable (or labeling) moiety (e.g., aradioactive, colorimetric, antigenic or enzymatic molecule, a detectablebead (such as a magnetic or electrodense (e.g., gold) bead), or amolecule that binds to another molecule (e.g., biotin or streptavidin),a therapeutic or diagnostic moiety (e.g., a radioactive, cytotoxic, orpharmaceutically active moiety), or a molecule that increases thesuitability of the antibody for a particular use (e.g., administrationto a subject, such as a human subject, or other in vivo or in vitrouses). Examples of molecules that can be used to derivatize an antibodyinclude albumin (e.g., human serum albumin) and polyethylene glycol(PEG). Albumin-linked and PEGylated derivatives of antibodies can beprepared using techniques well known in the art. In one embodiment, theantibody is conjugated or otherwise linked to transthyretin (TTR) or aTTR variant. The TTR or TTR variant can be chemically modified with, forexample, a chemical selected from the group consisting of dextran,poly(n-vinyl pyrrolidone), polyethylene glycols, propropylene glycolhomopolymers, polypropylene oxide/ethylene oxide co-polymers,polyoxyethylated polyols and polyvinyl alcohols.

Oligomers that contain one or more antigen binding proteins may beemployed as CD137 antagonists. Oligomers may be in the form ofcovalently-linked or non-covalently-linked dimers, trimers, or higheroligomers. Oligomers comprising two or more antigen binding protein arecontemplated for use, with one example being a homodimer. Otheroligomers include heterodimers, homotrimers, heterotrimers,homotetramers, heterotetramers, etc.

One embodiment is directed to oligomers comprising multiple antigenbinding proteins joined via covalent or non-covalent interactionsbetween peptide moieties fused to the antigen binding proteins. Suchpeptides may be peptide linkers (spacers), or peptides that have theproperty of promoting oligomerization. Leucine zippers and certainpolypeptides derived from antibodies are among the peptides that canpromote oligomerization of antigen binding proteins attached thereto, asdescribed in more detail below.

In particular embodiments, the oligomers comprise from two to fourantigen binding proteins. The antigen binding proteins of the oligomermay be in any form, such as any of the forms described above, e.g.,variants or fragments. Preferably, the oligomers comprise antigenbinding proteins that have CD137 binding activity.

In one embodiment, an oligomer is prepared using polypeptides derivedfrom immunoglobulins. Preparation of Fusion Proteins Comprising CertainHeterologous Polypeptides Fused to Various Portions of antibody-derivedpolypeptides (including the Fc domain) has been described, e.g., byAshkenazi et al., 1991, Proc. Natl. Acad. Sci. USA 88:10535; Byrn etal., 1990, Nature 344:677; and Hollenbaugh et al., 1992 “Construction ofImmunoglobulin Fusion Proteins”, in Current Protocols in Immunology,Suppl. 4, pages 10.19.1-10.19.11.

One embodiment is directed to a dimer comprising two fusion proteinscreated by fusing a CD137 binding fragment of an anti-CD137 antibody tothe Fc region of an antibody. The dimer can be made by, for example,inserting a gene fusion encoding the fusion protein into an appropriateexpression vector, expressing the gene fusion in host cells transformedwith the recombinant expression vector, and allowing the expressedfusion protein to assemble much like antibody molecules, whereuponinterchain disulfide bonds form between the Fc moieties to yield thedimer.

Another method for preparing oligomeric antigen binding proteinsinvolves use of a leucine zipper. Leucine zipper domains are peptidesthat promote oligomerization of the proteins in which they are found.Leucine zippers were originally identified in several DNA-bindingproteins (Landschulz et al., 1988, Science 240:1759), and have sincebeen found in a variety of different proteins. Among the known leucinezippers are naturally occurring peptides and derivatives thereof thatdimerize or trimerize. Examples of leucine zipper domains suitable forproducing soluble oligomeric proteins are described in WO 94/10308, andthe leucine zipper derived from lung surfactant protein D (SPD)described in Hoppe et al., 1994, FEBS Letters 344:191. The use of amodified leucine zipper that allows for stable trimerization of aheterologous protein fused thereto is described in Fanslow et al., 1994,Semin. Immunol. 6:267-78. In one approach, recombinant fusion proteinscomprising an anti-CD137 antibody fragment or derivative fused to aleucine zipper peptide are expressed in suitable host cells, and thesoluble oligomeric anti-CD137 antibody fragments or derivatives thatform are recovered from the culture supernatant.

Antigen binding proteins directed against CD137 can be used, forexample, in assays to detect the presence of CD137 polypeptides, eitherin vitro or in vivo. The antigen binding proteins also may be employedin purifying CD137 proteins by immunoaffinity chromatography. Suchantigen binding proteins that function as CD137 agonists may be employedin treating any CD137-induced condition, including but not limited tovarious cancers.

Antigen binding proteins may be employed in an in vitro procedure, oradministered in vivo to enhance CD137-induced biological activity.Disorders that would benefit (directly or indirectly) from activation ofCD137, examples of which are provided herein, thus may be treated. Inone embodiment, the present invention provides a therapeutic methodcomprising in vivo administration of a CD137 activating antigen bindingprotein to a mammal in need thereof in an amount effective forincreasing a CD137-induced biological activity.

In certain embodiments of the invention, antigen binding proteinsinclude fully human monoclonal antibodies that enhance a biologicalactivity of CD137.

Antigen binding proteins, including antibodies and antibody fragmentsdescribed herein, may be prepared by any of a number of conventionaltechniques. For example, they may be purified from cells that naturallyexpress them (e.g., an antibody can be purified from a hybridoma thatproduces it), or produced in recombinant expression systems, using anytechnique known in the art. See, for example, Monoclonal Antibodies,Hybridomas: A New Dimension in Biological Analyses, Kennet et al.(eds.), Plenum Press, New York (1980); and Antibodies: A LaboratoryManual, Harlow and Land (eds.), Cold Spring Harbor Laboratory Press,Cold Spring Harbor, N.Y., (1988).

Any expression system known in the art can be used to make therecombinant polypeptides, including antibodies and antibody fragmentsdescribed herein, of the invention. In general, host cells aretransformed with a recombinant expression vector that comprises DNAencoding a desired polypeptide. Among the host cells that may beemployed are prokaryotes, yeast or higher eukaryotic cells. Prokaryotesinclude gram negative or gram positive organisms, for example E. coli orbacilli. Higher eukaryotic cells include insect cells and establishedcell lines of mammalian origin. Examples of suitable mammalian host celllines include the COS-7 line of monkey kidney cells (ATCC CRL 1651)(Gluzman et al., 1981, Cell 23:175), L cells, 293 cells, C127 cells, 3T3cells (ATCC CCL 163), Chinese hamster ovary (CHO) cells, HeLa cells, BHK(ATCC CRL 10) cell lines, and the CV1/EBNA cell line derived from theAfrican green monkey kidney cell line CV1 (ATCC CCL 70) as described byMcMahan et al., 1991, EMBO J. 10: 2821. Appropriate cloning andexpression vectors for use with bacterial, fungal, yeast, and mammaliancellular hosts are described by Pouwels et al. (Cloning Vectors: ALaboratory Manual, Elsevier, N.Y., 1985).

The transformed cells can be cultured under conditions that promoteexpression of the polypeptide, and the polypeptide recovered byconventional protein purification procedures. One such purificationprocedure includes the use of affinity chromatography, e.g., over amatrix having all or a portion (e.g., the extracellular domain) of CD137bound thereto. Polypeptides contemplated for use herein includesubstantially homogeneous recombinant mammalian anti-CD137 antibodypolypeptides substantially free of contaminating endogenous materials.

Antigen binding proteins may be prepared, and screened for desiredproperties, by any of a number of known techniques. Certain of thetechniques involve isolating a nucleic acid encoding a polypeptide chain(or portion thereof) of an antigen binding protein of interest (e.g., ananti-CD137 antibody), and manipulating the nucleic acid throughrecombinant DNA technology. The nucleic acid may be fused to anothernucleic acid of interest, or altered (e.g., by mutagenesis or otherconventional techniques) to add, delete, or substitute one or more aminoacid residues, for example.

Polypeptides of the present disclosure can be produced using anystandard methods known in the art. In one example, the polypeptides areproduced by recombinant DNA methods by inserting a nucleic acid sequence(e.g., a cDNA) encoding the polypeptide into a recombinant expressionvector and expressing the DNA sequence under conditions promotingexpression.

Nucleic acids encoding any of the various polypeptides disclosed hereinmay be synthesized chemically. Codon usage may be selected so as toimprove expression in a cell. Such codon usage will depend on the celltype selected. Specialized codon usage patterns have been developed forE. coli and other bacteria, as well as mammalian cells, plant cells,yeast cells and insect cells. See for example: Mayfield et al., Proc.Natl. Acad. Sci. USA. 2003 100(2):438-42; Sinclair et al. Protein Expr.Purif. 2002 (1):96-105; Connell N D. Curr. Opin. Biotechnol. 200112(5):446-9; Makrides et al. Microbiol. Rev. 1996 60(3):512-38; andSharp et al. Yeast. 1991 7(7):657-78.

General techniques for nucleic acid manipulation are described forexample in Sambrook et al., Molecular Cloning: A Laboratory Manual,Vols. 1-3, Cold Spring Harbor Laboratory Press, 2 ed., 1989, or F.Ausubel et al., Current Protocols in Molecular Biology (Green Publishingand Wiley-Interscience: New York, 1987) and periodic updates, hereinincorporated by reference. The DNA encoding the polypeptide is operablylinked to suitable transcriptional or translational regulatory elementsderived from mammalian, viral, or insect genes. Such regulatory elementsinclude a transcriptional promoter, an optional operator sequence tocontrol transcription, a sequence encoding suitable mRNA ribosomalbinding sites, and sequences that control the termination oftranscription and translation. The ability to replicate in a host,usually conferred by an origin of replication, and a selection gene tofacilitate recognition of transformants is additionally incorporated.

The recombinant DNA can also include any type of protein tag sequencethat may be useful for purifying the protein. Examples of protein tagsinclude but are not limited to a histidine tag, a FLAG tag, a myc tag,an HA tag, or a GST tag. Appropriate cloning and expression vectors foruse with bacterial, fungal, yeast, and mammalian cellular hosts can befound in Cloning Vectors: A Laboratory Manual, (Elsevier, N.Y., 1985).

The expression construct is introduced into the host cell using a methodappropriate to the host cell. A variety of methods for introducingnucleic acids into host cells are known in the art, including, but notlimited to, electroporation; transfection employing calcium chloride,rubidium chloride, calcium phosphate, DEAE-dextran, or other substances;microprojectile bombardment; lipofection; and infection (where thevector is an infectious agent). Suitable host cells include prokaryotes,yeast, mammalian cells, or bacterial cells.

Suitable bacteria include gram negative or gram positive organisms, forexample, E. coli or Bacillus spp. Yeast, preferably from theSaccharomyces species, such as S. cerevisiae, may also be used forproduction of polypeptides. Various mammalian or insect cell culturesystems can also be employed to express recombinant proteins.Baculovirus systems for production of heterologous proteins in insectcells are reviewed by Luckow and Summers, (Bio/Technology, 6:47, 1988).Examples of suitable mammalian host cell lines include endothelialcells, COS-7 monkey kidney cells, CV-1, L cells, C127, 3T3, Chinesehamster ovary (CHO), human embryonic kidney cells, HeLa, 293, 293T, andBHK cell lines. Purified polypeptides are prepared by culturing suitablehost/vector systems to express the recombinant proteins. For manyapplications, the small size of many of the polypeptides disclosedherein would make expression in E. coli as the preferred method forexpression. The protein is then purified from culture media or cellextracts.

Proteins disclosed herein can also be produced using cell-translationsystems. For such purposes the nucleic acids encoding the polypeptidemust be modified to allow in vitro transcription to produce mRNA and toallow cell-free translation of the mRNA in the particular cell-freesystem being utilized (eukaryotic such as a mammalian or yeast cell-freetranslation system or prokaryotic such as a bacterial cell-freetranslation system. CD137-binding polypeptides can also be produced bychemical synthesis (e.g., by the methods described in Solid PhasePeptide Synthesis, 2nd ed., 1984, The Pierce Chemical Co., Rockford,Ill.). Modifications to the protein can also be produced by chemicalsynthesis.

The polypeptides of the present disclosure can be purified byisolation/purification methods for proteins generally known in the fieldof protein chemistry. Non-limiting examples include extraction,recrystallization, salting out (e.g., with ammonium sulfate or sodiumsulfate), centrifugation, dialysis, ultrafiltration, adsorptionchromatography, ion exchange chromatography, hydrophobic chromatography,normal phase chromatography, reversed-phase chromatography, gelfiltration, gel permeation chromatography, affinity chromatography,electrophoresis, countercurrent distribution or any combinations ofthese. After purification, polypeptides may be exchanged into differentbuffers and/or concentrated by any of a variety of methods known to theart, including, but not limited to, filtration and dialysis.

The purified polypeptide is preferably at least 85% pure, morepreferably at least 95% pure, and most preferably at least 98% pure.Regardless of the exact numerical value of the purity, the polypeptideis sufficiently pure for use as a pharmaceutical product.

In certain embodiments, the present disclosure provides monoclonalantibodies that bind to CD137. Monoclonal antibodies may be producedusing any technique known in the art, e.g., by immortalizing spleencells harvested from the transgenic animal after completion of theimmunization schedule. The spleen cells can be immortalized using anytechnique known in the art, e.g., by fusing them with myeloma cells toproduce hybridomas. Myeloma cells for use in hybridoma-producing fusionprocedures preferably are non-antibody-producing, have high fusionefficiency, and enzyme deficiencies that render them incapable ofgrowing in certain selective media which support the growth of only thedesired fused cells (hybridomas). Examples of suitable cell lines foruse in mouse fusions include Sp-20, P3-X63/Ag8, P3-X63-Ag8.653, NS1/1.Ag4 1, Sp210-Ag14, FO, NSO/U, MPC-11, MPC11-X45-GTG 1.7 and S194/5XX0 Bu1;examples of cell lines used in rat fusions include R210.RCY3, Y3-Ag1.2.3, IR983F and 48210. Other cell lines useful for cell fusions areU-266, GM1500-GRG2, LICR-LON-HMy2 and UC729-6.

Fragments or analogs of antibodies can be readily prepared by those ofordinary skill in the art following the teachings of this specificationand using techniques known in the art. Preferred amino- andcarboxy-termini of fragments or analogs occur near boundaries offunctional domains. Structural and functional domains can be identifiedby comparison of the nucleotide and/or amino acid sequence data topublic or proprietary sequence databases. Computerized comparisonmethods can be used to identify sequence motifs or predicted proteinconformation domains that occur in other proteins of known structureand/or function. Methods to identify protein sequences that fold into aknown three-dimensional structure are known. See, Bowie et al., 1991,Science 253:164.

Post-Translational Modifications of Polypeptides

In certain embodiments, the binding polypeptides of the invention mayfurther comprise post-translational modifications. Exemplarypost-translational protein modifications include phosphorylation,acetylation, methylation, ADP-ribosylation, ubiquitination,glycosylation, carbonylation, sumoylation, biotinylation or addition ofa polypeptide side chain or of a hydrophobic group. As a result, themodified soluble polypeptides may contain non-amino acid elements, suchas lipids, poly- or mono-saccharide, and phosphates. A preferred form ofglycosylation is sialylation, which conjugates one or more sialic acidmoieties to the polypeptide. Sialic acid moieties improve solubility andserum half-life while also reducing the possible immunogeneticity of theprotein. See Raju et al. Biochemistry. 2001 31; 40(30):8868-76.

In one embodiment, modified forms of the subject soluble polypeptidescomprise linking the subject soluble polypeptides to nonproteinaceouspolymers. In one embodiment, the polymer is polyethylene glycol (“PEG”),polypropylene glycol, or polyoxyalkylenes, in the manner as set forth inU.S. Pat. Nos. 4,640,835; 4,496,689; 4,301,144; 4,670,417; 4,791,192 or4,179,337.

PEG is a water soluble polymer that is commercially available or can beprepared by ring-opening polymerization of ethylene glycol according tomethods well known in the art (Sandler and Karo, Polymer Synthesis,Academic Press, New York, Vol. 3, pages 138-161). The term “PEG” is usedbroadly to encompass any polyethylene glycol molecule, without regard tosize or to modification at an end of the PEG, and can be represented bythe formula: X—O(CH₂CH₂O)_(n)—CH₂CH₂OH (1), where n is 20 to 2300 and Xis H or a terminal modification, e.g., a C₁₋₄ alkyl. In one embodiment,the PEG of the invention terminates on one end with hydroxy or methoxy,i.e., X is H or CH₃ (“methoxy PEG”). A PEG can contain further chemicalgroups which are necessary for binding reactions; which results from thechemical synthesis of the molecule; or which is a spacer for optimaldistance of parts of the molecule. In addition, such a PEG can consistof one or more PEG side-chains which are linked together. PEGs with morethan one PEG chain are called multiarmed or branched PEGs. Branched PEGscan be prepared, for example, by the addition of polyethylene oxide tovarious polyols, including glycerol, pentaerythriol, and sorbitol. Forexample, a four-armed branched PEG can be prepared from pentaerythrioland ethylene oxide. Branched PEG are described in, for example, EP-A 0473 084 and U.S. Pat. No. 5,932,462. One form of PEGs includes two PEGside-chains (PEG2) linked via the primary amino groups of a lysine(Monfardini et al., Bioconjugate Chem. 6 (1995) 62-69).

The serum clearance rate of PEG-modified polypeptide may be decreased byabout 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or even 90%, relative tothe clearance rate of the unmodified binding polypeptide. ThePEG-modified polypeptide may have a half-life (t_(1/2)) which isenhanced relative to the half-life of the unmodified protein. Thehalf-life of PEG-binding polypeptide may be enhanced by at least 10%,20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%,250%, 300%, 400% or 500%, or even by 1000% relative to the half-life ofthe unmodified binding polypeptide. In some embodiments, the proteinhalf-life is determined in vitro, such as in a buffered saline solutionor in serum. In other embodiments, the protein half-life is an in vivohalf life, such as the half-life of the protein in the serum or otherbodily fluid of an animal.

Therapeutic Methods, Formulations and Modes of Administration

The present disclosure further provides a method for treating a diseaserequiring either stimulation of immune responses, comprisingadministering an anti-CD137 polypeptide. Any of the antibodies disclosedherein may be used in such methods. For example, the methods may beperformed using an anti-CD137 polypeptide selected from the groupconsisting of a fully human antibody of an IgG class that binds to aCD137 epitope with a binding affinity of at least 10⁻⁶M, a Fab fullyhuman antibody fragment, having a variable domain region from a heavychain and a variable domain region from a light chain, a single chainhuman antibody, having a variable domain region from a heavy chain and avariable domain region from a light chain and a peptide linkerconnection the heavy chain and light chain variable domain regions,including the heavy and light chain variable regions (and CDRs withinsaid sequences) described in SEQ ID Nos. 1-143 (Table 5).

For example, in one embodiment, the methods disclosed herein include theuse of a fully human antibody having a heavy chain variable domainsequence that is at least 95% identical, at least 96% identical, atleast 97% identical, at least 98% identical, or at least 99% identical,to an amino acid sequences selected from the group consisting of SEQ IDNO. 1, SEQ ID NO. 3, SEQ ID NO. 5, SEQ ID NO. 6, SEQ ID NO. 7, SEQ IDNO. 9, SEQ ID NO. 11, SEQ ID NO. 13, SEQ ID NO. 15, SEQ ID NO. 17, SEQID NO. 19, SEQ ID NO. 21, SEQ ID NO. 23, SEQ ID NO. 25, SEQ ID NO. 27,SEQ ID NO. 29, SEQ ID NO. 31, SEQ ID NO. 33, SEQ ID NO. 35, SEQ ID NO.37, SEQ ID NO. 39, SEQ ID NO. 41, SEQ ID NO. 43, SEQ ID NO. 45, SEQ IDNO. 47, SEQ ID NO. 49, SEQ ID NO. 51, SEQ ID NO. 53, SEQ ID NO. 55, SEQID NO. 57, SEQ ID NO. 59, SEQ ID NO. 61, SEQ ID NO. 63, SEQ ID NO. 65,SEQ ID NO. 67, SEQ ID NO. 69, SEQ ID NO. 71, SEQ ID NO. 73, SEQ ID NO.75, SEQ ID NO. 77, SEQ ID NO. 79, SEQ ID NO. 81, SEQ ID NO. 83, SEQ IDNO. 85, SEQ ID NO. 87, SEQ ID NO. 89, SEQ ID NO. 91, SEQ ID NO. 93, SEQID NO. 95, SEQ ID NO. 97, SEQ ID NO. 99, SEQ ID NO. 101, SEQ ID NO. 103,SEQ ID NO. 105, SEQ ID NO. 107, SEQ ID NO. 109, SEQ ID NO. 111, SEQ IDNO. 113, SEQ ID NO. 115, SEQ ID NO. 117, SEQ ID NO. 119, SEQ ID NO. 121,SEQ ID NO. 123, SEQ ID NO. 125, SEQ ID NO. 127, SEQ ID NO. 129, SEQ IDNO. 130, SEQ ID NO. 131, SEQ ID NO. 132, SEQ ID NO. 133, SEQ ID NO. 134,SEQ ID NO. 135, SEQ ID NO. 136, SEQ ID NO. 137, SEQ ID NO. 138, SEQ IDNO. 139, SEQ ID NO. 140, SEQ ID NO. 141, SEQ ID NO. 142, and SEQ ID NO.143, and that having a light chain variable domain sequence that is atleast 95% identical, at least 96% identical, at least 97% identical, atleast 98% identical, or at least 99% identical, to an amino acidsequence selected from the group consisting of SEQ ID NO. 2, SEQ ID NO.4, SEQ ID NO. 6, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO.14, SEQ ID NO. 16, SEQ ID NO. 18, SEQ ID NO. 20, SEQ ID NO. 22, SEQ IDNO. 24, SEQ ID NO. 26, SEQ ID NO. 28, SEQ ID NO. 30, SEQ ID NO. 32, SEQID NO. 34, SEQ ID NO. 36, SEQ ID NO. 38, SEQ ID NO. 40, SEQ ID NO. 42,SEQ ID NO. 44, SEQ ID NO. 46, SEQ ID NO. 48, SEQ ID NO. 50, SEQ ID NO.52, SEQ ID NO. 54, SEQ ID NO. 56, SEQ ID NO. 58, SEQ ID NO. 60, SEQ IDNO. 62, SEQ ID NO. 64, SEQ ID NO. 66, SEQ ID NO. 68, SEQ ID NO. 70, SEQID NO. 72, SEQ ID NO. 74, SEQ ID NO. 76, SEQ ID NO. 78, SEQ ID NO. 80,SEQ ID NO. 82, SEQ ID NO. 84, SEQ ID NO. 86, SEQ ID NO. 88, SEQ ID NO.90, SEQ ID NO. 92, SEQ ID NO. 94, SEQ ID NO. 96, SEQ ID NO. 98, SEQ IDNO. 100, SEQ ID NO. 102, SEQ ID NO. 104, SEQ ID NO. 106, SEQ ID NO. 108,SEQ ID NO. 110, SEQ ID NO. 112, SEQ ID NO. 114, SEQ ID NO. 116, SEQ IDNO. 118, SEQ ID NO. 120, SEQ ID NO. 122, SEQ ID NO. 124, SEQ ID NO. 126,and SEQ ID NO. 128.

In one embodiment, the methods described herein include the use of afully human Fab antibody fragment has the heavy chain variable domainsequence that is at least 95% identical, at least 96% identical, atleast 97% identical, at least 98% identical, or at least 99% identical,to an amino acid sequences selected from the group consisting of SEQ IDNO. 1, SEQ ID NO. 3, SEQ ID NO. 5, SEQ ID NO. 6, SEQ ID NO. 7, SEQ IDNO. 9, SEQ ID NO. 11, SEQ ID NO. 13, SEQ ID NO. 15, SEQ ID NO. 17, SEQID NO. 19, SEQ ID NO. 21, SEQ ID NO. 23, SEQ ID NO. 25, SEQ ID NO. 27,SEQ ID NO. 29, SEQ ID NO. 31, SEQ ID NO. 33, SEQ ID NO. 35, SEQ ID NO.37, SEQ ID NO. 39, SEQ ID NO. 41, SEQ ID NO. 43, SEQ ID NO. 45, SEQ IDNO. 47, SEQ ID NO. 49, SEQ ID NO. 51, SEQ ID NO. 53, SEQ ID NO. 55, SEQID NO. 57, SEQ ID NO. 59, SEQ ID NO. 61, SEQ ID NO. 63, SEQ ID NO. 65,SEQ ID NO. 67, SEQ ID NO. 69, SEQ ID NO. 71, SEQ ID NO. 73, SEQ ID NO.75, SEQ ID NO. 77, SEQ ID NO. 79, SEQ ID NO. 81, SEQ ID NO. 83, SEQ IDNO. 85, SEQ ID NO. 87, SEQ ID NO. 89, SEQ ID NO. 91, SEQ ID NO. 93, SEQID NO. 95, SEQ ID NO. 97, SEQ ID NO. 99, SEQ ID NO. 101, SEQ ID NO. 103,SEQ ID NO. 105, SEQ ID NO. 107, SEQ ID NO. 109, SEQ ID NO. 111, SEQ IDNO. 113, SEQ ID NO. 115, SEQ ID NO. 117, SEQ ID NO. 119, SEQ ID NO. 121,SEQ ID NO. 123, SEQ ID NO. 125, SEQ ID NO. 127, SEQ ID NO. 129, SEQ IDNO. 130, SEQ ID NO. 131, SEQ ID NO. 132, SEQ ID NO. 133, SEQ ID NO. 134,SEQ ID NO. 135, SEQ ID NO. 136, SEQ ID NO. 137, SEQ ID NO. 138, SEQ IDNO. 139, SEQ ID NO. 140, SEQ ID NO. 141, SEQ ID NO. 142, and SEQ ID NO.143, and that has the light chain variable domain sequence that is atleast 95% identical, at least 96% identical, at least 97% identical, atleast 98% identical, or at least 99% identical, to the amino acidsequence consisting SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO.8, SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, SEQ ID NO. 16, SEQ IDNO. 18, SEQ ID NO. 20, SEQ ID NO. 22, SEQ ID NO. 24, SEQ ID NO. 26, SEQID NO. 28, SEQ ID NO. 30, SEQ ID NO. 32, SEQ ID NO. 34, SEQ ID NO. 36,SEQ ID NO. 38, SEQ ID NO. 40, SEQ ID NO. 42, SEQ ID NO. 44, SEQ ID NO.46, SEQ ID NO. 48, SEQ ID NO. 50, SEQ ID NO. 52, SEQ ID NO. 54, SEQ IDNO. 56, SEQ ID NO. 58, SEQ ID NO. 60, SEQ ID NO. 62, SEQ ID NO. 64, SEQID NO. 66, SEQ ID NO. 68, SEQ ID NO. 70, SEQ ID NO. 72, SEQ ID NO. 74,SEQ ID NO. 76, SEQ ID NO. 78, SEQ ID NO. 80, SEQ ID NO. 82, SEQ ID NO.84, SEQ ID NO. 86, SEQ ID NO. 88, SEQ ID NO. 90, SEQ ID NO. 92, SEQ IDNO. 94, SEQ ID NO. 96, SEQ ID NO. 98, SEQ ID NO. 100, SEQ ID NO. 102,SEQ ID NO. 104, SEQ ID NO. 106, SEQ ID NO. 108, SEQ ID NO. 110, SEQ IDNO. 112, SEQ ID NO. 114, SEQ ID NO. 116, SEQ ID NO. 118, SEQ ID NO. 120,SEQ ID NO. 122, SEQ ID NO. 124, SEQ ID NO. 126, and SEQ ID NO. 128.

In one embodiment, the methods described herein include the use of asingle chain human antibody having a heavy chain variable domainsequence that is at least 95% identical, at least 96% identical, atleast 97% identical, at least 98% identical, or at least 99% identicalto an amino acid sequence selected from the group consisting of SEQ IDNO. 1, SEQ ID NO. 3, SEQ ID NO. 5, SEQ ID NO. 6, SEQ ID NO. 7, SEQ IDNO. 9, SEQ ID NO. 11, SEQ ID NO. 13, SEQ ID NO. 15, SEQ ID NO. 17, SEQID NO. 19, SEQ ID NO. 21, SEQ ID NO. 23, SEQ ID NO. 25, SEQ ID NO. 27,SEQ ID NO. 29, SEQ ID NO. 31, SEQ ID NO. 33, SEQ ID NO. 35, SEQ ID NO.37, SEQ ID NO. 39, SEQ ID NO. 41, SEQ ID NO. 43, SEQ ID NO. 45, SEQ IDNO. 47, SEQ ID NO. 49, SEQ ID NO. 51, SEQ ID NO. 53, SEQ ID NO. 55, SEQID NO. 57, SEQ ID NO. 59, SEQ ID NO. 61, SEQ ID NO. 63, SEQ ID NO. 65,SEQ ID NO. 67, SEQ ID NO. 69, SEQ ID NO. 71, SEQ ID NO. 73, SEQ ID NO.75, SEQ ID NO. 77, SEQ ID NO. 79, SEQ ID NO. 81, SEQ ID NO. 83, SEQ IDNO. 85, SEQ ID NO. 87, SEQ ID NO. 89, SEQ ID NO. 91, SEQ ID NO. 93, SEQID NO. 95, SEQ ID NO. 97, SEQ ID NO. 99, SEQ ID NO. 101, SEQ ID NO. 103,SEQ ID NO. 105, SEQ ID NO. 107, SEQ ID NO. 109, SEQ ID NO. 111, SEQ IDNO. 113, SEQ ID NO. 115, SEQ ID NO. 117, SEQ ID NO. 119, SEQ ID NO. 121,SEQ ID NO. 123, SEQ ID NO. 125, SEQ ID NO. 127, SEQ ID NO. 129, SEQ IDNO. 130, SEQ ID NO. 131, SEQ ID NO. 132, SEQ ID NO. 133, SEQ ID NO. 134,SEQ ID NO. 135, SEQ ID NO. 136, SEQ ID NO. 137, SEQ ID NO. 138, SEQ IDNO. 139, SEQ ID NO. 140, SEQ ID NO. 141, SEQ ID NO. 142, and SEQ ID NO.143, and having a light chain variable domain sequence that is at least95% identical, at least 96% identical, at least 97% identical, at least98% identical, or at least 99% identical, to an amino acid sequenceconsisting of SEQ ID NO. 2, SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8,SEQ ID NO. 10, SEQ ID NO. 12, SEQ ID NO. 14, SEQ ID NO. 16, SEQ ID NO.18, SEQ ID NO. 20, SEQ ID NO. 22, SEQ ID NO. 24, SEQ ID NO. 26, SEQ IDNO. 28, SEQ ID NO. 30, SEQ ID NO. 32, SEQ ID NO. 34, SEQ ID NO. 36, SEQID NO. 38, SEQ ID NO. 40, SEQ ID NO. 42, SEQ ID NO. 44, SEQ ID NO. 46,SEQ ID NO. 48, SEQ ID NO. 50, SEQ ID NO. 52, SEQ ID NO. 54, SEQ ID NO.56, SEQ ID NO. 58, SEQ ID NO. 60, SEQ ID NO. 62, SEQ ID NO. 64, SEQ IDNO. 66, SEQ ID NO. 68, SEQ ID NO. 70, SEQ ID NO. 72, SEQ ID NO. 74, SEQID NO. 76, SEQ ID NO. 78, SEQ ID NO. 80, SEQ ID NO. 82, SEQ ID NO. 84,SEQ ID NO. 86, SEQ ID NO. 88, SEQ ID NO. 90, SEQ ID NO. 92, SEQ ID NO.94, SEQ ID NO. 96, SEQ ID NO. 98, SEQ ID NO. 100, SEQ ID NO. 102, SEQ IDNO. 104, SEQ ID NO. 106, SEQ ID NO. 108, SEQ ID NO. 110, SEQ ID NO. 112,SEQ ID NO. 114, SEQ ID NO. 116, SEQ ID NO. 118, SEQ ID NO. 120, SEQ IDNO. 122, SEQ ID NO. 124, SEQ ID NO. 126, and SEQ ID NO. 128.

In one embodiment, the fully human antibody has both a heavy chain and alight chain wherein the antibody has a heavy chain/light chain variabledomain sequence selected from the group consisting of SEQ ID NO. 1/SEQID NO. 2, SEQ ID NO. 3/SEQ ID NO. 4, SEQ ID NO. 5/SEQ ID NO. 6, SEQ IDNO. 7/SEQ ID NO. 8, SEQ ID NO. 9/SEQ ID NO. 10, SEQ ID NO. 11/SEQ ID NO.12, SEQ ID NO. 13/SEQ ID NO. 14, SEQ ID NO. 15/SEQ ID NO. 16, SEQ ID NO.17/SEQ ID NO. 18, SEQ ID NO. 19/SEQ ID NO. 20, SEQ ID NO. 21/SEQ ID NO.22, SEQ ID NO. 23/SEQ ID NO. 24, SEQ ID NO. 25/SEQ ID NO. 26, SEQ ID NO.27/SEQ ID NO. 28, SEQ ID NO. 29/SEQ ID NO. 30, SEQ ID NO. 31/SEQ ID NO.32, SEQ ID NO. 33/SEQ ID NO. 34, SEQ ID NO. 35/SEQ ID NO. 36, SEQ ID NO.37/SEQ ID NO. 38, SEQ ID NO. 39/SEQ ID NO. 40, SEQ ID NO. 41/SEQ ID NO.42, SEQ ID NO. 43/SEQ ID NO. 44, SEQ ID NO. 45/SEQ ID NO. 46, SEQ ID NO.47/SEQ ID NO. 48, SEQ ID NO. 49/SEQ ID NO. 50, SEQ ID NO. 51/SEQ ID NO.52, SEQ ID NO. 53/SEQ ID NO. 54, SEQ ID NO. 55/SEQ ID NO. 56, SEQ ID NO.57/SEQ ID NO. 58, SEQ ID NO. 59/SEQ ID NO. 60, SEQ ID NO. 61/SEQ ID NO.62, SEQ ID NO. 63/SEQ ID NO. 64, SEQ ID NO. 65/SEQ ID NO. 66, SEQ ID NO.67/SEQ ID NO. 68, SEQ ID NO. 69/SEQ ID NO. 70, SEQ ID NO. 71/SEQ ID NO.72, SEQ ID NO. 73/SEQ ID NO. 74, SEQ ID NO. 75/SEQ ID NO. 76, SEQ ID NO.77/SEQ ID NO. 78, SEQ ID NO. 79/SEQ ID NO. 80, SEQ ID NO. 81/SEQ ID NO.82, SEQ ID NO. 83/SEQ ID NO. 84, SEQ ID NO. 85/SEQ ID NO. 86, SEQ ID NO.87/SEQ ID NO. 88, SEQ ID NO. 89/SEQ ID NO. 90, SEQ ID NO. 91/SEQ ID NO.92, SEQ ID NO. 93/SEQ ID NO. 94, SEQ ID NO. 95/SEQ ID NO. 96, SEQ ID NO.97/SEQ ID NO. 98, SEQ ID NO. 99/SEQ ID NO. 100, SEQ ID NO. 101/SEQ IDNO. 102, SEQ ID NO. 103/SEQ ID NO. 104, SEQ ID NO. 105/SEQ ID NO. 106,SEQ ID NO. 107/SEQ ID NO. 108, SEQ ID NO. 109/SEQ ID NO. 110, SEQ ID NO.111/SEQ ID NO. 112, SEQ ID NO. 113/SEQ ID NO. 114, SEQ ID NO. 115/SEQ IDNO. 116, SEQ ID NO. 117/SEQ ID NO. 118, SEQ ID NO. 119/SEQ ID NO. 120,SEQ ID NO. 121/SEQ ID NO. 122, SEQ ID NO. 123/SEQ ID NO. 124, SEQ ID NO.125/SEQ ID NO. 126, SEQ ID NO. 127/SEQ ID NO. 128, SEQ ID NO. 129/SEQ IDNO. 28, SEQ ID NO. 130/SEQ ID NO. 28, SEQ ID NO. 131/SEQ ID NO. 28, SEQID NO. 132/SEQ ID NO. 28, SEQ ID NO. 133/SEQ ID NO. 28, SEQ ID NO.134/SEQ ID NO. 28, SEQ ID NO. 135/SEQ ID NO. 28, SEQ ID NO. 136/SEQ IDNO. 28, SEQ ID NO. 137/SEQ ID NO. 28, SEQ ID NO. 138/SEQ ID NO. 28, SEQID NO. 139/SEQ ID NO. 28, SEQ ID NO. 140/SEQ ID NO. 28, SEQ ID NO.141/SEQ ID NO. 28, SEQ ID NO. 142/SEQ ID NO. 28, and SEQ ID NO. 143/SEQID NO. 28.

In one embodiment, the fully human antibody Fab fragment has both aheavy chain variable domain region and a light chain variable domainregion wherein the antibody has a heavy chain/light chain variabledomain sequence selected from the group consisting of SEQ ID NO. 1/SEQID NO. 2, SEQ ID NO. 3/SEQ ID NO. 4, SEQ ID NO. 5/SEQ ID NO. 6, SEQ IDNO. 7/SEQ ID NO. 8, SEQ ID NO. 9/SEQ ID NO. 10, SEQ ID NO. 11/SEQ ID NO.12, SEQ ID NO. 13/SEQ ID NO. 14, SEQ ID NO. 15/SEQ ID NO. 16, SEQ ID NO.17/SEQ ID NO. 18, SEQ ID NO. 19/SEQ ID NO. 20, SEQ ID NO. 21/SEQ ID NO.22, SEQ ID NO. 23/SEQ ID NO. 24, SEQ ID NO. 25/SEQ ID NO. 26, SEQ ID NO.27/SEQ ID NO. 28, SEQ ID NO. 29/SEQ ID NO. 30, SEQ ID NO. 31/SEQ ID NO.32, SEQ ID NO. 33/SEQ ID NO. 34, SEQ ID NO. 35/SEQ ID NO. 36, SEQ ID NO.37/SEQ ID NO. 38, SEQ ID NO. 39/SEQ ID NO. 40, SEQ ID NO. 41/SEQ ID NO.42, SEQ ID NO. 43/SEQ ID NO. 44, SEQ ID NO. 45/SEQ ID NO. 46, SEQ ID NO.47/SEQ ID NO. 48, SEQ ID NO. 49/SEQ ID NO. 50, SEQ ID NO. 51/SEQ ID NO.52, SEQ ID NO. 53/SEQ ID NO. 54, SEQ ID NO. 55/SEQ ID NO. 56, SEQ ID NO.57/SEQ ID NO. 58, SEQ ID NO. 59/SEQ ID NO. 60, SEQ ID NO. 61/SEQ ID NO.62, SEQ ID NO. 63/SEQ ID NO. 64, SEQ ID NO. 65/SEQ ID NO. 66, SEQ ID NO.67/SEQ ID NO. 68, SEQ ID NO. 69/SEQ ID NO. 70, SEQ ID NO. 71/SEQ ID NO.72, SEQ ID NO. 73/SEQ ID NO. 74, SEQ ID NO. 75/SEQ ID NO. 76, SEQ ID NO.77/SEQ ID NO. 78, SEQ ID NO. 79/SEQ ID NO. 80, SEQ ID NO. 81/SEQ ID NO.82, SEQ ID NO. 83/SEQ ID NO. 84, SEQ ID NO. 85/SEQ ID NO. 86, SEQ ID NO.87/SEQ ID NO. 88, SEQ ID NO. 89/SEQ ID NO. 90, SEQ ID NO. 91/SEQ ID NO.92, SEQ ID NO. 93/SEQ ID NO. 94, SEQ ID NO. 95/SEQ ID NO. 96, SEQ ID NO.97/SEQ ID NO. 98, SEQ ID NO. 99/SEQ ID NO. 100, SEQ ID NO. 101/SEQ IDNO. 102, SEQ ID NO. 103/SEQ ID NO. 104, SEQ ID NO. 105/SEQ ID NO. 106,SEQ ID NO. 107/SEQ ID NO. 108, SEQ ID NO. 109/SEQ ID NO. 110, SEQ ID NO.111/SEQ ID NO. 112, SEQ ID NO. 113/SEQ ID NO. 114, SEQ ID NO. 115/SEQ IDNO. 116, SEQ ID NO. 117/SEQ ID NO. 118, SEQ ID NO. 119/SEQ ID NO. 120,SEQ ID NO. 121/SEQ ID NO. 122, SEQ ID NO. 123/SEQ ID NO. 124, SEQ ID NO.125/SEQ ID NO. 126, SEQ ID NO. 127/SEQ ID NO. 128, SEQ ID NO. 129/SEQ IDNO. 28, SEQ ID NO. 130/SEQ ID NO. 28, SEQ ID NO. 131/SEQ ID NO. 28, SEQID NO. 132/SEQ ID NO. 28, SEQ ID NO. 133/SEQ ID NO. 28, SEQ ID NO.134/SEQ ID NO. 28, SEQ ID NO. 135/SEQ ID NO. 28, SEQ ID NO. 136/SEQ IDNO. 28, SEQ ID NO. 137/SEQ ID NO. 28, SEQ ID NO. 138/SEQ ID NO. 28, SEQID NO. 139/SEQ ID NO. 28, SEQ ID NO. 140/SEQ ID NO. 28, SEQ ID NO.141/SEQ ID NO. 28, SEQ ID NO. 142/SEQ ID NO. 28, and SEQ ID NO. 143/SEQID NO. 28.

In one embodiment, the fully human single chain antibody has both aheavy chain variable domain region and a light chain variable domainregion, wherein the single chain fully human antibody has a heavychain/light chain variable domain sequence selected from the groupconsisting of SEQ ID NO. 1/SEQ ID NO. 2, SEQ ID NO. 3/SEQ ID NO. 4, SEQID NO. 5/SEQ ID NO. 6, SEQ ID NO. 7/SEQ ID NO. 8, SEQ ID NO. 9/SEQ IDNO. 10, SEQ ID NO. 11/SEQ ID NO. 12, SEQ ID NO. 13/SEQ ID NO. 14, SEQ IDNO. 15/SEQ ID NO. 16, SEQ ID NO. 17/SEQ ID NO. 18, SEQ ID NO. 19/SEQ IDNO. 20, SEQ ID NO. 21/SEQ ID NO. 22, SEQ ID NO. 23/SEQ ID NO. 24, SEQ IDNO. 25/SEQ ID NO. 26, SEQ ID NO. 27/SEQ ID NO. 28, SEQ ID NO. 29/SEQ IDNO. 30, SEQ ID NO. 31/SEQ ID NO. 32, SEQ ID NO. 33/SEQ ID NO. 34, SEQ IDNO. 35/SEQ ID NO. 36, SEQ ID NO. 37/SEQ ID NO. 38, SEQ ID NO. 39/SEQ IDNO. 40, SEQ ID NO. 41/SEQ ID NO. 42, SEQ ID NO. 43/SEQ ID NO. 44, SEQ IDNO. 45/SEQ ID NO. 46, SEQ ID NO. 47/SEQ ID NO. 48, SEQ ID NO. 49/SEQ IDNO. 50, SEQ ID NO. 51/SEQ ID NO. 52, SEQ ID NO. 53/SEQ ID NO. 54, SEQ IDNO. 55/SEQ ID NO. 56, SEQ ID NO. 57/SEQ ID NO. 58, SEQ ID NO. 59/SEQ IDNO. 60, SEQ ID NO. 61/SEQ ID NO. 62, SEQ ID NO. 63/SEQ ID NO. 64, SEQ IDNO. 65/SEQ ID NO. 66, SEQ ID NO. 67/SEQ ID NO. 68, SEQ ID NO. 69/SEQ IDNO. 70, SEQ ID NO. 71/SEQ ID NO. 72, SEQ ID NO. 73/SEQ ID NO. 74, SEQ IDNO. 75/SEQ ID NO. 76, SEQ ID NO. 77/SEQ ID NO. 78, SEQ ID NO. 79/SEQ IDNO. 80, SEQ ID NO. 81/SEQ ID NO. 82, SEQ ID NO. 83/SEQ ID NO. 84, SEQ IDNO. 85/SEQ ID NO. 86, SEQ ID NO. 87/SEQ ID NO. 88, SEQ ID NO. 89/SEQ IDNO. 90, SEQ ID NO. 91/SEQ ID NO. 92, SEQ ID NO. 93/SEQ ID NO. 94, SEQ IDNO. 95/SEQ ID NO. 96, SEQ ID NO. 97/SEQ ID NO. 98, SEQ ID NO. 99/SEQ IDNO. 100, SEQ ID NO. 101/SEQ ID NO. 102, SEQ ID NO. 103/SEQ ID NO. 104,SEQ ID NO. 105/SEQ ID NO. 106, SEQ ID NO. 107/SEQ ID NO. 108, SEQ ID NO.109/SEQ ID NO. 110, SEQ ID NO. 111/SEQ ID NO. 112, SEQ ID NO. 113/SEQ IDNO. 114, SEQ ID NO. 115/SEQ ID NO. 116, SEQ ID NO. 117/SEQ ID NO. 118,SEQ ID NO. 119/SEQ ID NO. 120, SEQ ID NO. 121/SEQ ID NO. 122, SEQ ID NO.123/SEQ ID NO. 124, SEQ ID NO. 125/SEQ ID NO. 126, SEQ ID NO. 127/SEQ IDNO. 128, SEQ ID NO. 129/SEQ ID NO. 28, SEQ ID NO. 130/SEQ ID NO. 28, SEQID NO. 131/SEQ ID NO. 28, SEQ ID NO. 132/SEQ ID NO. 28, SEQ ID NO.133/SEQ ID NO. 28, SEQ ID NO. 134/SEQ ID NO. 28, SEQ ID NO. 135/SEQ IDNO. 28, SEQ ID NO. 136/SEQ ID NO. 28, SEQ ID NO. 137/SEQ ID NO. 28, SEQID NO. 138/SEQ ID NO. 28, SEQ ID NO. 139/SEQ ID NO. 28, SEQ ID NO.140/SEQ ID NO. 28, SEQ ID NO. 141/SEQ ID NO. 28, SEQ ID NO. 142/SEQ IDNO. 28, and SEQ ID NO. 143/SEQ ID NO. 28.

In one embodiment, the anti-CD137 antibodies and antibody fragments ofthe invention are used to treat a disease is selected from the groupconsisting of cancers, autoimmune diseases and viral infections.

Due to the role of CD137 signaling in promoting T cell and NK cellproliferation, IFN-γ secretion, and prolonging the survival of CD8+ Tcells, CD137 engagement may provide an attractive strategy forimmunotherapy of cancer. Antibodies against CD137 have variableanti-tumor therapeutic effects depending on the immunogenicity of theexperimental tumor and anatomical site of tumor growth. Treatment withagonist anti-CD137 mAbs caused regression of large, well-establishedtumors in mice, including Ag104A sarcoma, P815 mastocytoma, EL4E7lymphomas and B10.2 fibrosarcoma. This treatment also generated systemicantitumor effects in established intracranial tumors including MCAsarcoma and GL261 glioma, but not in established subcutaneous andpulmonary tumors.

Altogether, CD137 stimulation results in enhanced expansion, survival,and effector functions of newly primed CD8+ T-cells, acting, in part,directly on these cells. Both CD4+ and CD8+ T-cells have been shown torespond to CD137 stimulation, however, it appears that enhancement ofT-cell function is greater in CD8+ cells (W. Shuford et al., J. Exp.Med., 186(1):47-55 (1997); I. Gramaglia et al., Eur. J. Immunol.,30(2):392-402 (2000); C. Takahashi et al., J. Immunol., 162:5037(1999)). Based on the critical role of CD137 stimulation in CD8+ T-cellfunction and survival, manipulation of the CD137/CD137L system providesan approach for the treatment of tumors and viral pathogens. Thus, inone embodiment, the anti-CD137 agonist antibodies of the invention(e.g., those described in Table 5) may be used in a method of treating apatient having a cancer, including, for example, ovarian cancer,colorectal cancer (e.g. colorectal adenocarcinoma), melanoma,hepatocellular carcinoma, renal cancer, breast cancer, head and neckcancer, lung cancer, non-hodgkin lymphoma, and liver cancer.

CD137 seems to play a role in CD8+ T cell-mediated antiviral responses.CD137L-deficient mice had decreased CTL responses to influenza virus inthe late stage of primary response and defective secondary response.There was also diminished CD8+ T cell responses and IFN-γ expressionafter lymphocytic choriomeningitis virus (LCMV) infection. There wasimpaired efficacy of vaccination with LCMV peptide in long termprotection generation against LCMV infection. CD137-deficient miceshowed decreased CTL activity against vesicular stomatitis virus (VSV).However, these mice showed normal humoral immune responses to viruses.

CD137 stimulation also restored CD8+ T cell response to animmunodominant influenza epitope in the absence of CD28 stimulation.Promoting CD8+ T cell responses by modifying CD137 signaling may be auseful approach to improve antiviral CD8+ T cell responses.

CD137/CD137L interaction plays an important role in regulatingalloresponses in vivo. Anti-CD137 mAbs enhance cardiac allograft andMHC-mismatched skin transplant rejection with dramatically increasedINF-γ production by CD8+ T cells and CTL activity against alloantigens.Blocking CD137/CD137L interaction by anti-CD137L mAbs significantlyinhibited rejection of intestinal allografts by CD8+ but not CD4+ Tcells. Anti-CD137 mAbs promoted both CD8+ and CD4+ T cell-mediatedgraft-versus-host disease (GVHD) and host anti-donor-mediated graftrejection could be regulated through CD137/CD137L interaction by usinganti-CD137 mAbs, CD137−/− donor T cells, or CD137L−/− recipients.Blocking CD137/CD137L interaction may reduce GVHD and prevent CD8+ Tcell-mediated allograft rejection. For allograft rejection involvingboth CD4+ and CD8+ T cells, combined blockade of CD137/CD137L and othercostimulatory signaling is required.

T cells are involved in the pathogenesis of many autoimmune diseases.The activation of T cells in response to their cognate peptide/MHCtargets requires costimulatory signals delivered by APCs occurring atmultiple steps. Conventional costimulation blockade is an attractivetherapeutic approach for the treatment of T cell-dependent auto immunediseases. Anti-CD137 Mabs can block several costimulatory pathways, suchas CD28/B7, CD40L/CD40 and OX-40L/OX-40R, with either soluble receptorsor neutralizing anti-ligand mAbs. However, costimulatory agonists ofCD137 could also prevent and have therapeutic effects on CD4+Tcell-involved autoimmune diseases. A single low dose of agonisticanti-CD137 mAb treatment prevented the development of EAE, a Th1cell-mediated demyelinating disease of the central nervous system usedas a murine model for human multiple sclerosis. Draining lymph nodecells from anti-4-1BB-treated mice failed to respond to antigenstimulation in vitro or to transfer disease to RAG-1-deficient recipientmice. When treatment was initiated after disease onset, early EAErelapse was also inhibited. Agonistic anti-4-1BB mAbs treatmentinitially increased T cell activation, and then promoted the clearanceof these activated CD4+ T cells, resulting in the attenuation of theireffector functions. Administering agonistic anti-CD137 mAbs also showedpromising therapeutic effect in both CD4+ T cell and B cell involvedspontaneous systemic autoimmune disease. MRL/lpr mice spontaneouslydevelop lymphadenopathy and a severe autoimmune disease resembling humanSLE due to the lymphoproliferative (lpr) mutation in the Fas gene.Short-term treatment with anti-CD137 blocked lymphadenopathy andspontaneous autoimmune diseases in MRL/lpr mice, ultimately leading totheir prolonged survival. This therapeutic regimen was also effectivewhen started after the mice had already showed clinically detectableautoimmune disease. The therapeutic effects of anti-CD137 were mediatedby the depletion of auto-reactive B cells, activated CD4+ T cells andthe aberrant CD4-CD8-B220+ CD3+ T cells that principally contribute tolymphadenopathy in MRL/lpr mice. Giving lupus-prone NZB×NZW F1 femalemice three injections of anti-CD137 mAbs between 26 and 35 weeks of agereversed acute disease, blocked chronic disease, and prolonged themice's lifespan. Autoantibody production in treated mice, regardless oftheir age or disease status, was rapidly suppressed without inducingsystemic immunosuppression or massive depletion of lymphocytes. In thismodel, adoptive transfer of antigen-primed CD4+ T cells or DCs overrodeanti-CD137-mediated protection, which suggests that unresponsiveness isnot achieved by active suppression. However, CD137 engagement in vivodoes not ameliorate all autoimmune diseases. Transgenic non-obesediabetic (NOD) mice overexpressing membrane-bound agonistic anti-CD137scFv in pancreatic beta cells exhibited increased GAD-specific T cellresponses, and developed more severe diabetes than their non-transgeniclittermates, with earlier onset, faster diabetic processes, and highermortality. Anti-CD137 treatment, starting around the onset of disease,promoted disease onset in NOD mice. Thus, in one embodiment, theanti-CD137 agonist antibodies of the invention (e.g., those described inTable 5) may be used in a method of treating a patient having anautoimmune disease, including, for example, multiple sclerosis,rheumatoid arthritis, systemic lupus erythematosus, and myaestheniagravis.

The present disclosure features methods for treating or preventing theS. aureus infection comprising administering an anti-CD137 polypeptide.Techniques and dosages for administration vary depending on the type ofspecific polypeptide and the specific condition being treated but can bereadily determined by the skilled artisan. In general, regulatoryagencies require that a protein reagent to be used as a therapeutic isformulated so as to have acceptably low levels of pyrogens. Accordingly,therapeutic formulations will generally be distinguished from otherformulations in that they are substantially pyrogen free, or at leastcontain no more than acceptable levels of pyrogen as determined by theappropriate regulatory agency (e.g., FDA).

Therapeutic compositions of the present disclosure may be administeredwith a pharmaceutically acceptable diluent, carrier, or excipient, inunit dosage form. Administration may be parenteral (e.g., intravenous,subcutaneous), oral, or topical, as non-limiting examples. In addition,any gene therapy technique, using nucleic acids encoding thepolypeptides of the invention, may be employed, such as naked DNAdelivery, recombinant genes and vectors, cell-based delivery, includingex vivo manipulation of patients' cells, and the like.

The composition can be in the form of a pill, tablet, capsule, liquid,or sustained release tablet for oral administration; or a liquid forintravenous, subcutaneous or parenteral administration; gel, lotion,ointment, cream, or a polymer or other sustained release vehicle forlocal administration.

In certain embodiments, the disclosed antibodies are administered byinhalation, but aerosolization of full IgG antibodies may prove limitingdue to their molecular size (˜150 kDa). To maximize available commercialaerosolization devices, smaller Fab fragments may be required. In thiscase, we may also need to generate Fab fragments from the parental IgGmolecules. Therefore, we will perform initial studies using standardenzyme-based digestion methodologies for the generation of Fabfragments, which will then be characterized in parallel with full IgGmolecules.

Methods well known in the art for making formulations are found, forexample, in “Remington: The Science and Practice of Pharmacy” (20th ed.,ed. A. R. Gennaro A R., 2000, Lippincott Williams & Wilkins,Philadelphia, Pa.). Formulations for parenteral administration may, forexample, contain excipients, sterile water, saline, polyalkylene glycolssuch as polyethylene glycol, oils of vegetable origin, or hydrogenatednapthalenes. Biocompatible, biodegradable lactide polymer,lactide/glycolide copolymer, or polyoxyethylene-polyoxypropylenecopolymers may be used to control the release of the compounds.Nanoparticulate formulations (e.g., biodegradable nanoparticles, solidlipid nanoparticles, liposomes) may be used to control thebiodistribution of the compounds. Other potentially useful parenteraldelivery systems include ethylene-vinyl acetate copolymer particles,osmotic pumps, implantable infusion systems, and liposomes. Theconcentration of the compound in the formulation varies depending upon anumber of factors, including the dosage of the drug to be administered,and the route of administration.

The polypeptide may be optionally administered as a pharmaceuticallyacceptable salt, such as non-toxic acid addition salts or metalcomplexes that are commonly used in the pharmaceutical industry.Examples of acid addition salts include organic acids such as acetic,lactic, pamoic, maleic, citric, malic, ascorbic, succinic, benzoic,palmitic, suberic, salicylic, tartaric, methanesulfonic,toluenesulfonic, or trifluoroacetic acids or the like; polymeric acidssuch as tannic acid, carboxymethyl cellulose, or the like; and inorganicacid such as hydrochloric acid, hydrobromic acid, sulfuric acidphosphoric acid, or the like. Metal complexes include zinc, iron, andthe like. In one example, the polypeptide is formulated in the presenceof sodium acetate to increase thermal stability.

Formulations for oral use include tablets containing the activeingredient(s) in a mixture with non-toxic pharmaceutically acceptableexcipients. These excipients may be, for example, inert diluents orfillers (e.g., sucrose and sorbitol), lubricating agents, glidants, andanti-adhesives (e.g., magnesium stearate, zinc stearate, stearic acid,silicas, hydrogenated vegetable oils, or talc).

Formulations for oral use may also be provided as chewable tablets, oras hard gelatin capsules wherein the active ingredient is mixed with aninert solid diluent, or as soft gelatin capsules wherein the activeingredient is mixed with water or an oil medium.

A therapeutically effective dose refers to a dose that produces thetherapeutic effects for which it is administered. The exact dose willdepend on the disorder to be treated, and may be ascertained by oneskilled in the art using known techniques. In general, the polypeptideis administered at about 0.01 μg/kg to about 50 mg/kg per day,preferably 0.01 mg/kg to about 30 mg/kg per day, most preferably 0.1mg/kg to about 20 mg/kg per day. The polypeptide may be given daily(e.g., once, twice, three times, or four times daily) or preferably lessfrequently (e.g., weekly, every two weeks, every three weeks, monthly,or quarterly). In addition, as is known in the art, adjustments for ageas well as the body weight, general health, sex, diet, time ofadministration, drug interaction, and the severity of the disease may benecessary, and will be ascertainable with routine experimentation bythose skilled in the art.

A CD137 binding polypeptide, as disclosed herein, can be administeredalone or in combination with one or more additional therapies such aschemotherapy radiotherapy, immunotherapy, surgical intervention, or anycombination of these. Long-term therapy is equally possible as isadjuvant therapy in the context of other treatment strategies, asdescribed above.

In certain embodiments of such methods, one or more polypeptidetherapeutic agents can be administered, together (simultaneously) or atdifferent times (sequentially). In addition, polypeptide therapeuticagents can be administered with another type of compounds for treatingcancer or for inhibiting angiogenesis.

In certain embodiments, the subject anti-CD137 antibodies agents of theinvention can be used alone.

In certain embodiments, the binding polypeptides of fragments thereofcan be labeled or unlabeled for diagnostic purposes. Typically,diagnostic assays entail detecting the formation of a complex resultingfrom the binding of a binding polypeptide to CD137. The bindingpolypeptides or fragments can be directly labeled, similar toantibodies. A variety of labels can be employed, including, but notlimited to, radionuclides, fluorescers, enzymes, enzyme substrates,enzyme cofactors, enzyme inhibitors and ligands (e.g., biotin, haptens).Numerous appropriate immunoassays are known to the skilled artisan (see,for example, U.S. Pat. Nos. 3,817,827; 3,850,752; 3,901,654; and4,098,876). When unlabeled, the binding polypeptides can be used inassays, such as agglutination assays. Unlabeled binding polypeptides canalso be used in combination with another (one or more) suitable reagentwhich can be used to detect the binding polypeptide, such as a labeledantibody reactive with the binding polypeptide or other suitable reagent(e.g., labeled protein A).

In one embodiment, the binding polypeptides of the present invention canbe utilized in enzyme immunoassays, wherein the subject polypeptides areconjugated to an enzyme. When a biological sample comprising a CD137protein is combined with the subject binding polypeptides, bindingoccurs between the binding polypeptides and the CD137 protein. In oneembodiment, a sample containing cells expressing a CD137 protein (e.g.,endothelial cells) is combined with the subject antibodies, and bindingoccurs between the binding polypeptides and cells bearing a CD137protein recognized by the binding polypeptide. These bound cells can beseparated from unbound reagents and the presence of the bindingpolypeptide-enzyme conjugate specifically bound to the cells can bedetermined, for example, by contacting the sample with a substrate ofthe enzyme which produces a color or other detectable change when actedon by the enzyme. In another embodiment, the subject bindingpolypeptides can be unlabeled, and a second, labeled polypeptide (e.g.,an antibody) can be added which recognizes the subject bindingpolypeptide.

In certain aspects, kits for use in detecting the presence of a CD137protein in a biological sample can also be prepared. Such kits willinclude a CD137 binding polypeptide which binds to a CD137 protein orportion of said receptor, as well as one or more ancillary reagentssuitable for detecting the presence of a complex between the bindingpolypeptide and the receptor protein or portions thereof. Thepolypeptide compositions of the present invention can be provided inlyophilized form, either alone or in combination with additionalantibodies specific for other epitopes. The binding polypeptides and/orantibodies, which can be labeled or unlabeled, can be included in thekits with adjunct ingredients (e.g., buffers, such as Tris, phosphateand carbonate, stabilizers, excipients, biocides and/or inert proteins,e.g., bovine serum albumin). For example, the binding polypeptidesand/or antibodies can be provided as a lyophilized mixture with theadjunct ingredients, or the adjunct ingredients can be separatelyprovided for combination by the user. Generally these adjunct materialswill be present in less than about 5% weight based on the amount ofactive binding polypeptide or antibody, and usually will be present in atotal amount of at least about 0.001% weight based on polypeptide orantibody concentration. Where a second antibody capable of binding tothe binding polypeptide is employed, such antibody can be provided inthe kit, for instance in a separate vial or container. The secondantibody, if present, is typically labeled, and can be formulated in ananalogous manner with the antibody formulations described above.

Polypeptide sequences are indicated using standard one- or three-letterabbreviations. Unless otherwise indicated, each polypeptide sequence hasamino termini at the left and a carboxy termini at the right; eachsingle-stranded nucleic acid sequence, and the top strand of eachdouble-stranded nucleic acid sequence, has a 5′ termini at the left anda 3′ termini at the right. A particular polypeptide sequence also can bedescribed by explaining how it differs from a reference sequence.

Having now described the present invention in detail, the same will bemore clearly understood by reference to the following examples, whichare included for purposes of illustration only and are not intended tobe limiting of the invention.

EXAMPLE 1

Human antibodies specific for human CD137 were identified and selectedfor therapeutic characteristics, including specificity for CD137 and ahigh degree of affinity for CD137 (e.g., at least 10⁻⁶M). The identifiedantibodies are described in Table 5.

To demonstrate agonistic activity, the ability of the anti-CD137antibodies (specifically D6, C3sh1G3, C7, and C3sh1F10) to mediatecostimulatory activity was assessed. Both anti-CD137 antibodies andanti-CD3 antibody were added to the wells of a 96 well plate in PBS toimmobilize the antibodies to the plate. The anti-CD137 was at 10microgram/ml and the anti-CD3 was at 3 microgram/ml. After a minimum of2 hours at room temperature, the wells were washed and monocyte depletedlymphocytes were added to the wells at 2×10⁵ per well. The monocyteswere depleted by labeling peripheral blood mononuclear cells (PBMC) withbiotinylated anti-CD14 antibody followed by incubation with anti-biotinmagnetic beads. Passage over a column in the presence of a magnetresulted in depletion of the monocytes.

To measure cell activation, the cells were labeled with FITC anti-humanCD25 after three days of culture. The percentage of cells positive forCD25 expression was measured by flow cytometry and is described in FIG.1A. The percent change from normal, or normalized data, was determinedby the below formula and the results are shown in FIG. 1B.

$\left( \frac{{{test}\mspace{14mu} \% \mspace{14mu} {CD}\; 25} - {{control}\mspace{14mu} \% \mspace{14mu} {CD}\; 25}}{{control}\mspace{20mu} \% \mspace{20mu} {CD}\; 25} \right) \times 100$

The use of immobilized anti-CD137 antibodies in the assay facilitatedtheir ability to mimic the ligand and promote signaling to the cellresulting in co-stimulation. From the data shown in FIG. 1A, antibody D6consistently, in particular, provided a co-stimulatory signal to the Tcells and is an agonistic anti-CD137 antibody. This is more apparentwhen the data are normalized relative to the control as shown in FIG.1B.

The above experiment was repeated in a third experiment testinganti-CD137 antibody D6 and D6 variants MA8, MB3, MSD2, MSH1, MSB7,MSA11, MD4, MB12, MB10, MB1, MSE5, MSC8, MSE3, MD1, and MC8 for theirability to increase CD25 activity as a measure of cell activation. Theexperiment was performed as described above, and the results areprovided in FIGS. 3A and 3B (normalized; for certain antibodies). Asshown in FIGS. 3A and 4B, out of many CD137 reactive antibodies tested,certain particular antibodies were identified that demonstrate agonisticactivity.

EXAMPLE 2

This example describes affinity characteristics for a number ofanti-CD137 antibodies that were identified and are described in Table 5.Table 1 describes the affinity characteristics of antibodies MC8 andMSA11. Table 2 describes the affinity characteristics of antibodiesMSB7, MSH1, and MD4. Table 3 describes the affinity characteristics ofantibodies D6, MB3, and MSC8. Table 4 describes the bindingcharacteristics of antibody B12.

Antibodies MC8, MSA1, MSB7, MSH1, MD4, D6, MB3, MSC8, and MB12 arevariants of the D6 antibody. Each antibody has a light chain variableregion comprising SEQ ID NO: 28, while the heavy chain of each is variedrelative to D6.

TABLE 1 Binding characteristics of antibodies MC8 and MSA11 Rmax name ka(1/Ms) kd (1/s) (RU) KA (1/M) KD (M) Chi2 MC8 2.08E5 6.28E−3 296 3.32E73.02E−8 0.807 MSA11 1.35E5 6.31E−3 293 2.13E7 4.69E−8 0.438

This example illustrates binding affinities of exemplary anti-CD137antibodies disclosed herein. Affinities were determined using surfaceplasmon resonance (Biacore). Briefly, anti-human Fc antibody (GE,BR-1008-39) was immobilized on CM5 sensor chip to approximately 1000 RUusing standard NHS/EDC coupling methodology. Antibodies (about 10 μg/ml)were captured for 60 s at a flow rate 10 μl/min Recombinant humanCD137/His was serially diluted in running buffer (HBS-EP). Allmeasurements were conducted with a flow rate of 30 μL/min. Surfaces wereregenerated with 3M MgCl₂ for 60 s. A 1:1 (Langmuir) binding model wasused to fit the data.

EXAMPLE 3

The following example describes the characterization of anti-CD137antibodies D6, MB3, MSC8, and MB12. The amino acid sequence of thevariable heavy and light chains of each of these antibodies is providedin Table 5.

Cross-reactivity studies of anti-CD137 antibodies D6, MB3, MSC8, andMB12 revealed that these antibodies are specific to human CD137 and tonot cross react with murine CD137. Results from the cross-reactivitystudy are described in FIG. 2 and show that each antibody was specificfor human CD137, and not murine CD137.

A Maxisorp plate was coated with recombinant human CD137/Fc or mouseCD137/Fc at 2 μg/mL at 4° C., overnight. The plate was blocked for 1hour at room temperature, washed 3 times with PBS-Tween (PBST), thenanti-CD137 antibodies (˜1 μg/mL) diluted in casein were added andincubated for 30 min with shaking. The plate was washed 3 times withPBST, horseradish peroxidase (HRP)-conjugated mouse anti-human Lambda(1:1000 in casein) was added, then 3,3′,5,5′-Tetramethylbenzidine (TMB)was added as substrate and developed about 5 min. 2M H₂SO₄ was used tostop the reaction and the OD was read at 450 nm.

TABLE 5  Heavy and Light Chain Variable Domain Amino Acid SequencesHeavy chain variable  Light chain variable  domain regionsdomain regions C3sh1A2 EVQLVQSGGGVVQPGRSLRLSCAASGAIQMTQSPSSLSASVGDRVSFTCQASQ FTFSSYAMHWVRQAPGKGLEWVAADISNYLNWYQQKPGKAPKLMISDAST MSHDGIQKDYADSVKGRFTISRDNSKLETGVPSRFSGSGSGTYFTFTISSLQPD NTLYLQMNSLRAEDTAVYYCAQGGGDFATYYCQHYDSFPLTFGGGTKVEIK FAYGMEDYWGQGTLVTVSS SEQ ID SEQ ID NO. 74NO. 73 C3sh1A5 QVQLVQSGAEEKTPGASVKISCKASG DIVMTQTPSSLSASVGDRVTITCRASQNTFNNYDIHWVRQAPGERPEWMGWI GIYNYLAWYQQKLGKAPNLLIYATSNNSGNGDTRNSQKFQGRVTITWDTSAS LQSGVPSRFSGSGSGTDFTLTISSLQPETAYMELSSLTSEDTGVYFCARAEGPL DFATYYCQQSYSTPWTFGQGTKVEIKDYWGQGTLVTVSS SEQ ID NO. 75 SEQ ID NO. 76 C3sh1A9QVQLVQSGAEVKKPGASVKVSCKAS SSELTQDPAVSVALGQTVRITCQGDSLGYTFTSKWMHWVRQAPGQGPEWMG RRYYASWYQQKPGQAPRLLIYGKNIRVINPSSGGTTYAQKFQGRLTVTRDTSS PSGIPDRFSGTDSGNTDFLTITGAQAESTVYMELSSLRSEDTAVYYCARDDVF DEADYYCNSRDTDANQPLVLFGGGTDYYFGLDVWGQGTTVTVSS SEQ ID KLTVL SEQ ID NO. 78 NO. 77 C3sh1B2QVQLVQSGGDLVQPGGSLRLSCAASG SYELTQPPSVSVSPGQTARITCSGEKLFLFSNSWMTWVRQAPGKGLEWLANI DDKYTFWYQQRTGQTPVLVIYQDKKKPDGSGQYYVDSLRGRFTISRDNAKN RPSGIPERFSGSNSGNTATLTISGTQAVSLYLQMNSLRVEDTAMYYCARDRGN DEADYYCQTYDSGAPVFGGGTKLTVDGLDYWGQGTLVTVSS SEQ ID  L SEQ ID NO. 80 NO. 79 C3sh1B4 EVQLVESGAELKKPGASVKVSCMAS QSVLTQPASVSGSPGQSITISCTGTSSDGYTFTDYYMHWVRQAPGQGLEWMG VGGYDFVAWYQQHPGKAPKLLIYDVWINPNSGGTNYAQKFQGRVTMTRDT SNRPSGVSNRFSGSKSGNTASLKISGLSISTAYMELSRLRSDDTAVYYCARDG RAEDEADYYCSSYTSSSARWVFGGGTSTYTDYWGQGTLVTVSS SEQ ID  KVTVL SEQ ID NO. 82 NO. 81 C3sh1B6QVQLVESGGGLVKPGRSLRLSCTASG SYELTQPPSVSVAPGKTARITCGGNNIFTFGDYAMSWFRQAPGKGLEWVGFI GSKSVHWYQQKPGQAPVLVIYYDSDRSKAFGGTTEYAASVKGRFTISRDDS RPSGIPERFSGSNSGNTATLTISRVEAGNSIAYLQMNSLKTEDTAVYYCTRDSG DEADYYCQVWDSSSDHPVFGGGTKLPGWERSFDYWGQGTLVTVSS SEQ ID TVL SEQ ID NO. 84 NO. 83 C3sh1B9QVQLVQSGAEEKTPGASVKISCKASG DIVMTQSPSSLSASVGDRVTITCRASQNTFNNYDIHWVRQAPGERPEWMGWI GIYNYLAWYQQKLGKAPNLLIYATSNNSGNGDTRNSQKFQGRVTITWDTSAS LQSGVPSRFSGSGSGTDFTLTISSLQPETAYMELSSLTSEDTGVYFCARAEGPL DFATYYCQQSYSTPWTFGQGTKVEIKDYWGQGTLVTVSS SEQ ID NO. 85 SEQ ID NO. 86 C3sh1C1EVQLVQSGAEVKKPGASVKVSCKAS SYELTQPPSVSVAPGKTARITCGGNNIGYTFTGYYMHWVRQAPGQSPEWMG GSKHVHWYQQKPGQAPVLVINYDSDWINVGNGNIRYSQKFQGRVTFTGDTS RPSGIPERLSGSNSGNTATLTISRVEAGATTAYMDLSSLRSEDTAVFYCAREGA DEADYYCQVWDSTSDHVIFGGGTKLASGLDLDYWGQGTLVTVSS SEQ ID TVL SEQ ID NO. 88 NO. 87 C3sh1C2EVQLLESGGGVVQPGRSLRLSCAASG QSVLTQPPSVSVAPGKTARITCGGNNIFTFSNYAMHWVRQAPGKGLEWVAVI GSKSVHWYQQKPGQAPVLVIYYDSDSLDGSNRHYADSVKGRFTISRDNSQN RPSGIPERFSGSNSGNTATLTISRVEAGTLYLQMNSLRAEDTAMYYCAQDLYD DEADYYCQVWDSSSDHVVFGGGTKLDNRWGVFDYWGQGTLVTVSS SEQ TVL SEQ ID NO. 90 ID NO. 89 C3sh1C7EVQLLESGAEVKKPGASVKVSCKASG SYVLTQPPSASGSPGQSVTISCTGTSSDYTFTSYYMHWVRQAPGQGLEWMGII VGGYNYVSWYQQHPGKAPKLMIYDNPSGGSTSYAQKFQGRVTMTRDTSTS VSKRPSGVPDRFSGSKSGNTASLTISGTVYMELSSLRSEDTAVYYCARDPGA LQAEDEADYYCSSYTSSSTRYVFGTGGGYFDYWGQGTLVTVSS SEQ ID  TKLTVL SEQ ID NO. 92 NO. 91 C3sh1D11 QVQLVQSGAEMKKPGSSVKVSCKAS QSALTQPPSVSAAPGQKVTISC SGSSSGYTFTSYGISWVRQAPGQGLEWMGW NIGNNYVSWYQQLPGTAPKLLIYDNNISAYNGNTNYAQKLQGRVTMTTDTST KRPSGIPDRFSGSKSGTSATLGITGLQTSTAYMELRSLRSDDTAVYYCARDLSQ GDEADYYCGTWDSSLSAVVFGGGTKWYQLYGADYYYGMDVWGQGTTVT VTVL SEQ ID NO. 94 VSS SEQ ID NO. 93 C3sh1D4EVQLVQSGAEVTKPGASVKVSCKAS QAGLTQPASVSVSPGQSITISCTGTSSDGYTFTGYYMHWVRQAPGQGLEWMG VGAYNYVSWYQQHPGKAPKLMIYDWINPNSGGTNYAQKFQGRVTMTRDT VSNRPSGVSNRFSGSKSGNTASLTISGSISTAYMELSRLRSDDTAVYYCARDN LQAEDEADYYCSSYSSINSRYVFGTGAGLGDYWGQGTLVTVSS SEQ ID  TKVTVL SEQ ID NO. 96 NO. 95 C3sh1D6EVQLVESGAEVKKPGSSVKVSCKASG QSVLTQPASVSGSPGQSITISCTGTSSDGTFSSYAISWVRQAPGQGLEWMGIIN VGGYNYVSWYQQYPGKAPKLMIYDPSGGSTSYAQKFQGRVTMTRDTSTST VSKRPSGVSHRFSGSKSGNTASLTISGVYMELSSLRSEDTAVYYCAGTPSLKY LQAEDEADYYCSSYTSSSTLVFGGGTDYYYYGMDVWGQGTTVTVSS SEQ KLTVL SEQ ID NO. 98 ID NO. 97 C3sh1E12EVQLVESGAEVKKPGASVKVSCKAS SSELTQDPAVSVALGQTVRITCQGDSLGYTFTNYYIHWVRQAPGQGLEWMGII RSYYASWYQQKPGQAPLLVIYGKNNNPSGGYTSSAQKFQGRVTMTRDTSTS RPAGISDRFSGSDSEDIASLTITGAQAETVYMELSSLRSEDTAVYYCARDRDSG DEADYYCNSRDSNAHWVFGGGTKLTSYYDAFDIWGQGTMVTVSS SEQ ID VL SEQ ID NO. 100 NO. 99 C3sh1E7QVQLVQSGGGLVQPGGSLRLSCAASG QSVVTQPPSVSAAPGHKVTISC SGNSSFTFSSSAMSWVRQAPGKGLEWVSGIS NVGRNYVSWYQQVPGTAPKLLIYDDGSGDSAYYADSVKGRFTISRDNSKNT NKRPSGIPDRFSGSTSGASATLVITGLLYLHMNSLTAEDTAVYYCASGGNYG QTGDEADYYCGAWDSSLSAGVFGGGSGTIVSHGMDVWGQGTTVTVSS SEQ TKLTVL SEQ ID NO. 102 ID NO. 101 C3sh1E9EVQLLESGGGLVQPGGSLRLSCAASG SYELTQPPSVSVSPGQTARITCSGDALFTFNNYAMSWVRQAPGKGLEWVSTI PKQYAYWYQQKPGQAPVLVIYKDSESGSGENTHYADSVKGRFTISRDNSKD RPSGIPERFSGSSSGTTVTLTISGVQAETLYLQMSSLRAEDTAVYYCANQYDT DEADYYCQSADSSGTYVVFGGGTKLTDYYYWGEYFHHWGQGTLVTVSS TVL SEQ ID NO. 104 SEQ ID NO. 103 C3sh1F1EVQLVESGAEVKKPGASVKVSCKAS QPVLTQPASVSGSPGQSITISCTGTSSDGYTFTGYYMHWVRQAPGQGLEWMG VGSYNFVSWYQQHPGKAPKLMIYDVWINPNSGGTNYAQKFQGRVTMTRDT SNRPSGVSDRFSGSKSGNTASLTISGLSTSTVYMELSSLRSEDTAVYYCARDI QAEDEADYYCSSYTSSSTRWVFGGGTRAFDIWGQGTMVTVSS SEQ ID  KLTVL SEQ ID NO. 106 NO. 105 C3sh1F10EVQLVQSGVEVKKPGASVKVSCKVS LPVLTQPPSVSGAPGQRVTISCTGSSSGNTLTEISMHWVRQVPGKGLEWMGG NIGAGYDVHWYQQLPGTAPKLLIYGFDLEDGETVYAQKFQGRVTLTEDTSI NSNRPSGVPDRFSGSKSGTSASLAITGDTAYELRSLRSEDTAVYYCATGPAGY LQAEDEADYYCQSYDSSLSGYVFGTGRLFEYWGQGTLVTVSS SEQ ID  TKVTVL SEQ ID NO. 108 NO. 107 C3sh1F12EVQLVQSGAEVKKPGASVKVSCKAS QSVLTQPPSASGTPGQRVTISCSGSSSGYTFTSHYMHWVRQAPGQGLEWMG NIGSNTVNWYQQLPGTAPKLLIYSNNVINPSGGSTSYAQKFQGRVTMTRDTS QRPSGVPDRFSGSKSGTSASLAISGLQTSTVYMDLSSLRSEDTAVYYCARRSE SEDEADYYCAAWDDSLNGWVFGGGAYYHGMDVWGQGTTVTVSS SEQ ID TKLTVL SEQ ID NO. 110 NO. 109 C3sh1F2EVQLVESGGGLVKPGESLRLSCAASG DIVMTQTPLSLPVTPGEPASISCRSSQSFTFKSYPMAWVRQAPGKGLEWVSSIS LLYSNGYNYLDWYLQKPGQSPQLLIYSSGDHRYYADSVKGRFTISRDNARNS WGSNRASGVPDRFSGSGSGTDFTLKISLSLQMNNLRAEDTAVYYCPAGRDFD RVEAEDVGIYYCMQALHVPPYTFGQHWGRGTLVTVSS SEQ ID NO. 111 GTKVEIK SEQ ID NO. 112 C3sh1G1QVQLVESGAEVKKPGASVKVSCKAS SYELTQPPSVSVAPGKTATITCGGDTIGYTFTDYYIHWVRQAPGQGLEWVG GSKVVHWYQQKPGQAPVLVMYYDSWINPNSGGTNYAQRFQGRVTMTRDT ERPSGIPERFSASNSGNTATLTISRVEASISTTYMELSRLRFDDTAVYYCASDP GDEADYYCQVWDSGSVVFGGGTKLTGGNPYFDYWGQGTLVTVSS SEQ ID VL SEQ ID NO. 114 NO. 113 C3sh1G11 EVQLVESGGGVVQPGRSLRLSCAASG SYELTQPPSASGTPGQRVTISCSGSSSNFTFSSYGMHWVRQAPGKGLEWVALI IGPYSINWYQQLPGTAPKLLIHSNTQRSYDGTNKYYADSVKGRFTISRDNSKN PSGVPDRFSGSKSGTSASLAISGLQSETLYLQMNSLSSEDTALYYCASNHDIL DEADYYCAAWDGSLNGVVFGGGTQTGGDYWGQGTLVTVSS SEQ ID NO. LTVL SEQ ID NO. 116 115 C3sh1G2EVQLVESGGALVQPGGSLRLSCAGSG DIVMTQTPSSLSASVGDRVTITCRASQFTFSNFWMHWVRQAPGKGLEWVADI SISSYLNWYQQKPGKAPKLLIYAASSLSGDGSEKYYVDSVKGRFTFSRDNARN QSGVPSRFSGSGSGTDFTLTISSLQPEDSLYLQMNSLRIEDTAVYYCARDAMR FATYYCQQSYSTPHFGGGTKVEIKGGDLDYWGQGTLVTVSS SEQ ID  SEQ ID NO. 118 NO. 117 C3sh1G3EVQLVQSGAEVKKPGSSVKVSCKASG QSVVTQPPSVSAAPGHKVTISCSGNSSGTFSSYAISWVRQAPGQGLEWMGGII NVGRNYVSWYQQVPGTAPKLLIYDDPIFGTANYAQKFQGRVTITADESTSTA NKRPSGIPDRFSGSQSGTSATLGITGLYMELSSLRSEDTAVYYCAGRFDYYDS QTGDEADYYCGTWDSSLTLYVFGTGSGYYYGPFDYWGQGTLVTVSS SEQ TKLTVL SEQ ID NO. 120 ID NO. 119 C3sh1G5EVQLVQSGAEVKKPGESLKISCKGSG QSVVTQPPSVSAAPGQKVTISCSGSDSYSFTSYWIGWVRQMPGKGLEWMGII NIGNNYVSWYQQVPGTAPKLLIYDNYYPGDSDTIYSPSFQGQVTLSADKSTST KRPSGIPDRFSGSKSGTSATLGITGLQTAYVQWNSLKASDTAVYYCARLTVSG GDEADYYCVTGDGGLGAVVFGGGTKSSTTTGGMDVWGHGTTVTVSS SEQ LTVL SEQ ID NO. 122 ID NO. 121 C3sh1G8QVQLVQSGAEVKKPGASVKVSCKTS SSELTQDPAVSVALGQTVRITCQGDSLGYNFNTYYIHWVRQAPGQGLEWMGI RRYYASWYQQKPGQAPLLVMFGEDKINPSGGYTSYAQNFQGRVTMTRDTST RPSGIPDRFSGSSSGNTASLTITGAQAESTVYMELSSLRSEDTALYYCARELGG DEADYYCNSRDTSGSWVFGGGTKLTNVRRDDAFDIWGQGTMVTVSS SEQ VL SEQ ID NO. 124 ID NO. 123 C3sh1H10QMQLVQSGAEVKKPGASVKVSCKAS QSVLTQPASVSGSPGQSITISCTGTSSDGYTFTSYYLHWVRQAPGQGLEWMGI VGGYNYVSWYQQHPGKAPKLMIYDIHSSGGSTTYAQKFQGRVTMTRDTST VSNRPSGVSNRFSGSKSGNTASLTISGSTVYMELSSLRSEDTAVYYCARGYFG LQAEDEADYYCSSYTSSSTYVFGTGTSGSFDYWGHGTLVTVSS SEQ ID  KLTVL SEQ ID NO. 126 NO. 125 C3sh1H4QVQLVQSGAEVKQPGASVKVSCKAS SYVLTQPASVSGSPGQSITISCTGTSSDGYTFTNNYMHWVRQAPGQGLEWMG VGGYNYVSWYQQHPGKAPKLMIYDIINPTGGSTTYAQKFQGRVIMTTDTST VSKRPSGVSNRFSGSKSGNTASLTISGSTVFMELSSLRSEDTAVYYCARDLGE LQAEDEADYYCSSYTSSSTLVFGTGTLLLAFDYWGQGTLVTVSS SEQ ID KVTVL SEQ ID NO. 128 NO. 127 MA8QMQLVQSGAEVKKPGASVKVSCKAS SYELTQPPSVSVAPGKTARITCGGNNIGYRFGGYYMHWVRQAPGQGLEWM GSKSVHWYQQKPGQAPVLVIYYDSDGWINPNSGGTNYAQKFQGRVTITRDT RPSGIPERFSGSNSGNTATLTISRVEAGSASTAYMELSSLRSEDTAVYYCAREG DEADYYCQVWDSSSVVFGGGTQLTVEAVGLDLDYWGQGTLVTVSS SEQ ID L SEQ ID NO. 28 NO. 129 MB1QMQLVQSGAEVKKPGASVKVSCKAS SYELTQPPSVSVAPGKTARITCGGNNIGYGMKGYYMHWVRQAPGQGLEWM GSKSVHWYQQKPGQAPVLVIYYDSDGWINPNSGGTNYAQKFQGRVTITRDT RPSGIPERFSGSNSGNTATLTISRVEAGSASTAYMELSSLRSEDTAVYYCAREG DEADYYCQVWDSSSVVFGGGTQLTVEAVGLDLDLWGQGTLVTVSS SEQ ID L SEQ ID NO. 28 NO. 130 MB3QMQLVQSGAEVKKPGASVKVSCKAS SYELTQPPSVSVAPGKTARITCGGNNIGYQMRGYYMHWVRQAPGQGLEWM GSKSVHWYQQKPGQAPVLVIYYDSDGWINPNSGGTNYAQKFQGRVTITRDT RPSGIPERFSGSNSGNTATLTISRVEAGSASTAYMELSSLRSEDTAVYYCAREG DEADYYCQVWDSSSVVFGGGTQLTVEAVGLDLDYWGQGTLVTVSS SEQ ID L SEQ ID NO. 28 NO. 131 MB10QMQLVQSGAEVKKPGASVKVSCKAS SYELTQPPSVSVAPGKTARITCGGNNIGYLMQGYYMHWVRQAPGQGLEWM GSKSVHWYQQKPGQAPVLVIYYDSDGWINPNSGGTNYAQKFQGRVTITRDT RPSGIPERFSGSNSGNTATLTISRVEAGSASTAYMELSSLRSEDTAVYYCAREG DEADYYCQVWDSSSVVFGGGTQLTVEAVGLDLDYWGQGTLVTVSS SEQ ID L SEQ ID NO. 28 NO. 132 MB12QMQLVQSGAEVKKPGASVKVSCKAS SYELTQPPSVSVAPGKTARITCGGNNIGYSLEGYYMHWVRQAPGQGLEWMG GSKSVHWYQQKPGQAPVLVIYYDSDWINPNSGGTNYAQKFQGRVTITRDTS RPSGIPERFSGSNSGNTATLTISRVEAGASTAYMELSSLRSEDTAVYYCAREGE DEADYYCQVWDSSSVVFGGGTQLTVAVGLDLDYWGQGTLVTVSS SEQ ID L SEQ ID NO. 28 NO. 133 MC8QMQLVQSGAEVKKPGASVKVSCKAS SYELTQPPSVSVAPGKTARITCGGNNIGYMPDGYYMHWVRQAPGQGLEWM GSKSVHWYQQKPGQAPVLVIYYDSDGWINPRTGGTNYAQKFQGRVTITRDT RPSGIPERFSGSNSGNTATLTISRVEAGSASTAYMELSSLRSEDTAVYYCAREG DEADYYCQVWDSSSVVFGGGTQLTVAAFRLELDAWGQGTLVTVSS SEQ ID L SEQ ID NO. 28 NO. 134 MD1QMQLVQSGAEVKKPGASVKVSCKAS SYELTQPPSVSVAPGKTARITCGGNNIGYRLQGYYMHWVRQAPGQGLEWM GSKSVHWYQQKPGQAPVLVIYYDSDGWINPNSGGTNYAQKFQGRVTITRDT RPSGIPERFSGSNSGNTATLTISRVEAGSASTAYMELSSLRSEDTAVYYCAREG DEADYYCQVWDSSSVVFGGGTQLTVEAVGLDLDYWGQGTLVTVSS SEQ ID L SEQ ID NO. 28 NO. 135 MD4QMQLVQSGAEVKKPGASVKVSCKAS SYELTQPPSVSVAPGKTARITCGGNNIGYNWTGYYMHWVRQAPGQGLEWM GSKSVHWYQQKPGQAPVLVIYYDSDGWINPMAGGTNYAQKFQGRVTITRD RPSGIPERFSGSNSGNTATLTISRVEAGTSASTAYMELSSLRSEDTAVYYCARE DEADYYCQVWDSSSVVFGGGTQLTVGWARGVELDMWGQGTLVTVSS SEQ L SEQ ID NO. 28 ID NO. 136 MSA11QMQLVQSGAEVKKPGASVKVSCKAS SYELTQPPSVSVAPGKTARITCGGNNIGYSFTGYYMHWVRQAPGQGLEWMG GSKSVHWYQQKPGQAPVLVIYYDSDWVNPKSGGTNYAQKFQGRVTITRDTS RPSGIPERFSGSNSGNTATLTISRVEAGASTAYMELSSLRSEDTAVYYCAREG DEADYYCQVWDSSSVVFGGGTQLTVWARRIDLDEWGQGTLVTVSS SEQ ID L SEQ ID NO. 28 NO. 137 MSB7QMQLVQSGAEVKKPGASVKVSCKAS SYELTQPPSVSVAPGKTARITCGGNNIGYSFSGYYMHWVRQAPGQGLEWMG GSKSVHWYQQKPGQAPVLVIYYDSDWVNPMSGGTNYAQKFQGRVTITRDT RPSGIPERFSGSNSGNTATLTISRVEAGSASTAYMELSSLRSEDTAVYYCAREG DEADYYCQVWDSSSVVFGGGTQLTVMAMRLELDKWGQGTLVTVSS SEQ L SEQ ID NO. 28 ID NO. 138 MSD2QMQLVQSGAEVKKPGASVKVSCKAS SYELTQPPSVSVAPGKTARITCGGNNIGYNFAGYYMHWVRQAPGQGLEWM GSKSVHWYQQKPGQAPVLVIYYDSDGWVNPQSGGTNYAQKFQGRVTITRD RPSGIPERFSGSNSGNTATLTISRVEAGTSASTAYMELSSLRSEDTAVYYCARE DEADYYCQVWDSSSVVFGGGTQLTVGEGRGLDLDWWGQGTLVTVSS SEQ L SEQ ID NO. 28 ID NO. 139 MSE3QMQLVQSGAEVKKPGASVKVSCKAS SYELTQPPSVSVAPGKTARITCGGNNIGYNFSGYYMHWVRQAPGQGLEWMG GSKSVHWYQQKPGQAPVLVIYYDSDWINPKSGGTNYAQKFQGRVTITRDTS RPSGIPERFSGSNSGNTATLTISRVEAGASTAYMELSSLRSEDTAVYYCAREGG DEADYYCQVWDSSSVVFGGGTQLTVARGVDLDTWGQATLVTVSS SEQ ID L SEQ ID NO. 28 NO. 140 MSE5QMQLVQSGAEVKKPGASVKVSCKAS SYELTQPPSVSVAPGKTARITCGGNNIGYSFGGYYMHWVRQAPGQGLEWMG GSKSVHWYQQKPGQAPVLVIYYDSDWVNPNSGGTNYAQKFQGRVTITRDTS RPSGIPERFSGSNSGNTATLTISRVEAGASTAYMELSSLRSEDTAVYYCAREGY DEADYYCQVWDSSSVVFGGGTQLTVGLGLDLDVWGQGTLVTVSS SEQ ID L SEQ ID NO. 28 NO. 141 MSC8QMQLVQSGAEVKKPGASVKVSCKAS SYELTQPPSVSVAPGKTARITCGGNNIGYNFGGYYMHWVRQAPGQGLEWM GSKSVHWYQQKPGQAPVLVIYYDSDGWVNPKSGGTNYAQKFQGRVTITRD RPSGIPERFSGSNSGNTATLTISRVEAGTSASTAYMELSSLRSEDTAVYYCARE DEADYYCQVWDSSSVVFGGGTQLTVGEAVGLDLDYWGQGTLVTVSS L SEQ ID NO. 28 SEQ ID NO. 142 MSH1QMQLVQSGAEVKKPGASVKVSCKAS SYELTQPPSVSVAPGKTARITCGGNNIGYNFGGYYMHWVRQAPGQGLEWM GSKSVHWYQQKPGQAPVLVIYYDSDGWVNPHSGGTNYAQKFQGRVTITRD RPSGIPERFSGSNSGNTATLTISRVEAGTSASTAYMELSSLRSEDTAVYYCARE DEADYYCQVWDSSSVVFGGGTQLTVGEAWGLDLDLWGQGTLVTVSS SEQ L SEQ ID NO. 28 ID NO. 143

INCORPORATION BY REFERENCE

The contents of all references, patents, pending patent applications andpublished patents, cited throughout this application are herebyexpressly incorporated by reference.

We claim:
 1. An isolated fully human anti-CD137 antibody of an IgG classthat binds to a CD137 epitope, comprising a heavy chain/light chainvariable domain sequence SEQ ID NO. 138/SEQ ID NO. 28 (called MSB7herein).
 2. An anti-CD137 fully human antibody Fab fragment or ananti-CD137 single chain human antibody, comprising a heavy chain/lightchain variable domain sequence SEQ ID NO. 138/SEQ ID NO. 28 (called MSB7herein).
 3. A method of treating cancer in a subject in need thereof,the method comprising administering an effective amount of the antibodyor antibody fragment of claims 1, such that the cancer is treated. 4.The method of claim 3, wherein the cancer is selected from the groupconsisting of: ovarian cancer, colorectal cancer, melanoma,hepatocellular carcinoma, renal cancer, breast cancer, head and neckcancer, lung cancer and liver cancer.
 5. The method of claim 3, whereinthe disease is selected from the group consisting of cancers, autoimmunediseases and viral infections.